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2-0523 March 2005

DRAFT ASSESSMENT REPORT

APPLICATION A544

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FOOD STANDARDS AUSTRALIA NE+ ,EALAND !FSAN,'

FSANZs role is to protect the health and safety of people in Australia and New Zealand through themaintenance of a safe food supply. FSANZ is a partnership between ten Go ernments: the AustralianGo ernment! Australian States and "erritories! and New Zealand. #t is a statutory authority under$ommonwealth law and is an independent% e&pert body.

FSANZ is responsible for de eloping% arying and re iewing standards and for de eloping codes ofconduct with industry for food a ailable in Australia and New Zealand co ering labelling%composition and contaminants. #n Australia% FSANZ also de elops food standards for food safety%ma&imum residue limits% primary production and processing and a range of other functions including

the coordination of national food sur eillance and recall systems% conducting research and assessing policies about imported food.

"he FSANZ 'oard appro es new standards or ariations to food standards in accordance with policyguidelines set by the Australia and New Zealand Food (egulation )inisterial $ouncil *)inisterial$ouncil+ made up of Australian Go ernment% State and "erritory and New Zealand ,ealth )inistersas lead )inisters% with representation from other portfolios. Appro ed standards are then notified tothe )inisterial $ouncil. "he )inisterial $ouncil may then re-uest that FSANZ re iew a proposed ore&isting standard. #f the )inisterial $ouncil does not re-uest that FSANZ re iew the draft standard%

or amends a draft standard% the standard is adopted by reference under the food laws of the AustralianGo ernment% States% "erritories and New Zealand. "he )inisterial $ouncil can% independently of anotification from FSANZ% re-uest that FSANZ re iew a standard.

"he process for amending the Australia New Zealand Food Standards Code is prescribed in the FoodStandards Australia New Zealand Act 1 1 *FSANZ Act+. "he diagram below represents thedifferent stages in the process including when periods of public consultation occur. "his process

aries for matters that are urgent or minor in significance or comple&ity.

INITIALComment on scope possiblef An IA report is prepared with an outline of issues and

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IN)ITATION FOR PUBLIC SUBMISSIONS

FSANZ has prepared a /raft Assessment (eport of Application A0 ! and prepared a draftariation to the Australia New Zealand Food Standards Code *the $ode+ !

FSANZ in ites public comment on this /raft Assessment (eport based on regulation impact principles and the draft ariation to the $ode for the purpose of preparing an amendment tothe $ode for appro al by the FSANZ 'oard.

2ritten submissions are in ited from interested indi iduals and organisations to assistFSANZ in preparing the Final Assessment for this Application. Submissions should% where possible% address the ob3ecti es of FSANZ as set out in section 14 of the FSANZ Act.#nformation pro iding details of potential costs and benefits of the proposed change to the$ode from sta5eholders is highly desirable. $laims made in submissions should be supportedwhere er possible by referencing or including rele ant studies% research findings% trials%sur eys etc. "echnical information should be in sufficient detail to allow independentscientific assessment.

"he processes of FSANZ are open to public scrutiny% and any submissions recei ed willordinarily be placed on the public register of FSANZ and made a ailable for inspection. #fyou wish any information contained in a submission to remain confidential to FSANZ% youshould clearly identify the sensiti e information and pro ide 3ustification for treating it ascommercial6in6confidence. Section 78 of the FSANZ Act re-uires FSANZ to treat in6confidence% trade secrets relating to food and any other information relating to food% thecommercial alue of which would be% or could reasonably be e&pected to be% destroyed or

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Assessment reports are a ailable for iewing and downloading from the FSANZ website.Alternati ely% re-uests for paper copies of reports or other general in-uiries can be directed toFSANZs #nformation

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CONTENTS

E=ECUTI)E SUMMAR> AND STATEMENT OF REASONS88888888888888888888888888888888888888888

# N"($"# A"

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ATTAC*MENT 3 - FOOD TEC*NOLOG> REPORT88888888888888888888888888888888888888888888888888883

ATTAC*MENT 4 - SAFET> ASSESSMENT REPORT88888888888888888888888888888888888888888888888888843ATTAC*MENT 5 - DIETAR> E=POSURE ASSESSMENT REPORT888888888888888888888888886

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E $c % ;$ S ar( a". S%a%$ $"% 9 R$a/ "/

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FSANZ recei ed an Application on 8 August 44 from >nile er Australia imited% toamend Standard 1.7.7 H rocessing Aids of the Australia New Zealand Food Standards Code *the $ode+ to appro e the use of #ce Structuring rotein "ype ### , $ 1 *#S + as a

processing aid for the preparation of ice cream and edible ices. "he Applicant re-uested thatthe term edible ices include froIen yoghurts and froIen fruit andJor egetable 3uices and

drin5s.

#ce structuring proteins are naturally occurring proteins and peptides that are found in aariety of li ing organisms such as fish% plants% insects% fungi and bacteria which protect them

from damage in ery cold conditions that would normally cause organisms to freeIe. Anumber of these products are consumed in foods so ice structuring proteins are a normalcomponent of the human diet.

For use in manufacturing ice cream and edible ices% ice structuring proteins do not actually pre ent freeIing but influence the growth and structure of ice crystal formation and hence the physical properties of froIen foods. For ice cream and edible ices these include hardness%thermal stability *including impro ed heat shoc5 resistance during storage andtransportation+% creaminess% mouth6feel% and fla our deli ery.

R$: 1a% r( pr #1$ a". # $c% ;$

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R / a//$// $"%

A number of criteria ha e been addressed in the safety assessment including: acharacterisation of the #S gene transferred to the production organism% its origin% functionand stability! a characterisation of the functional protein present in the #S preparationsecreted by the genetically modified yeast *G) yeast+! and the potential for the #S

preparation to be either to&ic or allergenic to humans.

No potential public health and safety concerns ha e been identified in the safety assessment

of #S .

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Submissions are now in ited on this report to assist FSANZ to complete the FinalAssessment.

S%a%$ $"% 9 R$a/ "/

"he draft ariation to Standard 1.7.7 H rocessing Aids of the $ode to permit the use of #Sas a processing aid for the manufacture of ice cream and edible ice products is recommendedfor the following reasons.

"he safety assessment concluded that no public health and safety concerns associatedwith using #S as a processing aid for the manufacture of ice cream and edible ice products ha e been identified.

"he use of #S is technologically 3ustified to alter the properties of ice cream and edible

ice products. #S binds to and influences the growth and structure of the de eloping icecrystals during manufacture% which alters the physical and sensory properties of thefinal products.

As concluded by the regulatory impact analysis% the costs that would arise from a

ariation to Standard 1.7.7 to permit the use of #S as a processing aid for themanufacture of ice cream and edible ice products do not outweigh the direct andindirect benefits to the community% Go ernment or industry that would arise from the

ariation.

"h d d f i i h $ d i i i h h i 14 b3 i f

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8 I"%r . c% "

FSANZ recei ed an Application on 8 August 44 from >nile er Australia imited% toamend Standard 1.7.7 H rocessing Aids of the Australia New Zealand Food Standards Code *the $ode+ to appro e the use of #ce Structuring rotein "ype ### , $ 1 *#S + as a

processing aid for the preparation of ice cream and edible ices. "he Applicant re-uested thatedible ices include froIen yoghurts and froIen fruit andJor egetable 3uices and drin5s. Forthis report% #S refers to the specific ice structuring protein of the Application and not ageneric class of proteins. "he Application is for the appro al of the specific #S product%

rather than appro al for the broad class of ice structuring proteins that may e&ist.

2or5 on this Group 7 *cost6reco ered+ Application commenced on 4 August 44 .

#ce structuring proteins are naturally occurring proteins and peptides that are found in aariety of li ing organisms such as fish% plants% insects% fungi and bacteria. "hese proteins

help to protect the organisms from damage in ery cold conditions that would normally causethem to freeIe. A number of these products are present in commonly consumed foods% so icestructuring proteins are already a natural component of the human diet.

"he Applicant wishes to use #S during the manufacture of froIen ice products. For use inmanufacturing ice cream and edible ices% ice structuring proteins do not actually pre entfreeIing but influence the growth and structure of ice crystal formation and hence physicaland sensory properties of froIen foods. roperties rele ant for froIen ice products includethermal stability% hardness% creaminess% mouth6feel and fla our deli ery.

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or processing% but does not perform a technological function in thefinal food! and

*b+ the substance is used in the course of manufacture of a food at thelowest le el necessary to achie e a function in the processing ofthat food% irrespecti e of any ma&imum permitted le el specified.

#S is a processing aid and not a food additi e for the purposes of this Application since itfulfils its technological purpose during the manufacture of the froIen ice products and doesnot perform a technological function of a food additi e in the final food. #S does not fitneatly into any of the possible food additi e functions listed in Schedule 0 of Standard 1.7.1 H Food Additi es.

#S 9binds to and influences the growth and structure of the de eloping ice crystals during production of such products. "his different ice structure alters the properties of the food products. "he altered properties of the ice structure are not due to the presence of #S byitself% but the effect #S has on the ice structure formation during processing. Stability of iced

products containing #S is due to the ice structure that has been formed% rather than theresidual presence of #S .

38 O# $c% ;$

"he ob3ecti e of this assessment is to determine whether it is appropriate to amend the $odeto permit the use of #S as a processing aid for the manufacture of ice cream and edible ices.

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any written policy guidelines formulated by the )inisterial $ouncil.

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48 Bac :r ".

48 * /% r ca1 Bac :r ".

Darious naturally occurring proteins and peptides ha e been e&tracted and identified from the blood of fish li ing in ery cold water. "hese proteins and peptides protect the fish from thedamage that would be caused by freeIing and allow them to sur i e and were identified o erthirty years ago. Similar proteins were subse-uently also found in many other organisms thatsur i e in ery cold en ironments% such as plants% insects% fungi and bacteria. A number of

these proteins are already consumed in foods that ha e been significant parts of the humandiet% such as fish and carrots.

"hese proteins ha e been 5nown as thermal hysteresis proteins or antifreeIe proteins.,owe er% since they do not pre ent ice forming but modify the structure and growth of icecrystals they ha e been gi en the name 9ice structuring proteins.

#ce structuring proteins affect the growth and structure of ice crystals by directly 9binding *or more correctly 9adsorbing or 9accumulating+ to the growing ice crystals and inhibiting thegrowth *particularly in one direction+ resulting in modification of the resulting ice structureand hence its physical properties. "he mechanisms of the binding to ice crystals for differenttypes of ice structuring proteins has been postulated by arious groups to include hydrogen

bonding% and hydrophobic and hydrophilic interactions. (egardless of how the proteinswor5% their addition during manufacture causes changes to ice crystal siIe and structurewhich also alters the ices physical properties. For food products based on ice% addition of#S also has important impacts on the sensory properties of the resultant ice products. Such

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0.1.1. ,istory of >se

,umans ha e pre iously been e&posed to ice structuring proteins in the diet through theconsumption of certain fish and egetable species. #S is present in the blood of ocean pout%a species of cold6water fish found off the northeast coast of North America% that is har estedcommercially for human food.

Food6grade yeasts are used widely in the manufacture of beer% wine% and for production ofenIymes including those used in cheese manufacture. "he production organism for #S is

ba5ers yeast * Sacc&arom#ces cerevisiae + which has a long history of safe use in thelea ening of bread.

0.1.1.7 /escription of the Genetic )odification

"he gene encoding #S is a synthetic ersion of the gene deri ed from ocean pout. "he genefrom ocean pout was resynthesised to impro e production and secretion of the protein in the

production organism *ba5ers yeast+. "he synthetic gene in yeast encodes the identical amino

acid se-uence to that of the nati e #S deri ed from ocean pout. "he gene cassette did notcontain any antibiotic resistance mar5er genes or any bacterial /NA.

"!1!1!% C&aracterisation of ISP

#S type ### , $ 1 consists of a 5nown se-uence of amino acids% and studies on its properties and the physical structure of the protein ha e been published. 'iochemicalanalysis of the yeast6deri ed #S confirms that the protein is the same as the nati e #S from

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#n studies using human olunteers% ingestion of #S preparation for eight wee5s at a highdaily dose did not result in specific antibody formation% indicating that #S is not li5ely to beany more immunogenic than the ma3ority of dietary proteins.

Additional biochemical analyses simulating gastric fluid digestion with pepsin in an in vitro test system showed that both #S and its inacti e form would be readily degraded in thehuman digesti e system. #n addition% amino acid se-uence analysis showed a susceptibilityto proteolytic brea5down by intestinal enIymes such as trypsin. "hese results indicate that#S is therefore unli5ely to be absorbed intact or accumulate in the body. 'ased on a thorough assessment of allergic potential% and the results of the analytical% animal%human% andin vitro data presented in this application% #S preparation is not to&ic and isunli5ely to e o5e an allergic reaction in fish6sensitised indi iduals% or to sensitise potentiallysusceptible indi iduals in the wider population.

0.1.1. $onclusion

No potential public health and safety concerns ha e been identified in the assessment of #S .

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7 mgJday for toddlers!

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7 mgJday for primary school children aged 061 years! and 8 mgJday for teenagers aged 17618 years.

"he 80 th percentile dietary e&posures for consumers of #S for New Zealand were estimatedas:

mgJday for the whole population aged 10 years and abo e! and 7 mgJday for teenagers aged 10618 years.

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Secondly% the #S preparation is a protein6rich mi&ture that has been shown to be readilydegraded in the gastrointestinal system% as e&pected of normal dietary protein.

Finally% gi en the a ailable data on #S *chemical% biochemical% to&icological andallergenicity+% the intended low le el of use% and its acceptable bac5ground in food% its use asa processing aid in froIen products such as ice cream does not raise any safety concerns.

582 Na% r$ 9 %h$ c$ /%r c% r ": pr %$ "

"he ice structuring protein of this Application was originally found in a cold water fish% ocean pout% 2acro3oarces americanus4 found along the North American coast. "he ocean pout isconsumed by humans as fish% although the Application states that current stoc5s are o er6fished.

"he Application states that the serum of the ocean pout contains at least 1 different types ofice structuring proteins which can be separated by high performance li-uid chromatography*, $+. "he protein of this Application is one of these 1 proteins which has been separated

and purified and which the Applicant calls #S type ### , $ 1 . "his protein is the mostabundant and has the most acti e functionality from in vitro ice6structuring tests. #t is madeup of amino acids in a 5nown se-uence with a molecular weight of appro&imately ? 5/a."he protein is heat tolerant% with an isoelectric point between H 14% is stable between p, H 1 and is not glycocon3ugated *that is the protein is not bound with carbohydrates+.

"he identified #S was selected for commercial production due to its good functionality andthermal and p, stability. "he Application states that it was considered not economic or

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583 Pr p /$. 9 . /$

"he Applicant proposes to use #S to alter the properties of a number of ice creams and edibleice products% some of which may be new or uni-ue compared to those that are currentlya ailable or possible with present technology and ingredients. "he Applicant has stated the

products they wish to use #S for are those contained *standardised+ under item 7 H #ce creamand edible ices in Schedule 1 of Standard 1.7.1 H Food Additi es. "he Applicant re-uestedthat such items include ice creams% froIen yoghurts and froIen fruit andJor egetable 3uicesand drin5s.

As discussed abo e% #S 9binds to and influences the growth and structure of the de elopingice crystals during production of such products. "his different ice structure alters the

properties of the food products. According to the Applicant% one important ad antage is thatthe froIen ice products ha e impro ed resistance to melting which is a ma3or ad antageagainst temperature abuse and also allows the de elopment of inno ati e new products. Aswell the ice crystal structure is altered which offers impro ed sensory deli ery of fla oursand colours. "hat is% fla ours and colours are not so easily drawn out of the ice crystal

structure by a consumer of a froIen ice product% as the new altered ice structure impedes thisand allows for more e en distribution.

"he Applicant states another possible ad antage that the use of #S offers is the commercial production of new inno ati e products with consumer benefits. Such new products includeconsumer acceptable lowJIero fat products% products with higher fruit content and productswith low added sugar content. "he altered ice structure pro ides opportunities to de elop

products due to the altered physical properties% te&ture and mouth6feel.

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#S has also been appro ed for use in ,ong Eong% )e&ico% the hilippines and #ndonesia.$ommercial product is sold in the >SA and the hilippines. "he Applicant is also applying

for appro al in a number of other countries. "he Applicant states that where appro al has been sought% no re3ections ha e been made.

585 La#$11 ": // $/

"here are a number of rele ant labelling issues for this Application% which could arise fromregulating #S as a processing aid. "hese include consideration of labelling for processingaids which are produced using gene technology and labelling for foods deri ed fromsubstances that may cause ad erse reactions.

"he following sections outline the rele ant labelling issues for the different aspects of thisApplication.

"!"!1 Processin- aid

$lause 7 *d+ of Standard 1. . H abelling of #ngredients% e&empts processing aids fromingredient labelling re-uirements. ,owe er% there are other possible labelling re-uirementsrele ant for this Application that must be considered. $lause of Standard 1. .7 H)andatory 2arning and Ad isory Statements and /eclarations% re-uires the labelling ofsubstances that may cause ad erse reactions to food *see section 0.0. +. Standard 1.0. HFood roduced using Gene "echnology re-uires labelling for processing aids or foodadditi es produced using gene technology if the food contains no el /NA andJor no el

proteins *see section 0.0.7+.

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Fish and fish products)il5 and mil5 products

eanuts and soybeans% and their productsAdded Sulphites in concentrations of 14 mgJ5g or more"ree nuts and sesame seeds and their products

As #S is not a fish or fish product% but is produced from yeast there is no re-uirement tolabel under the re-uirements of $lause of Standard 1. .7. "he Applicant also asserts thatthere are no allergenicity concerns with #S % although it is identical to a protein from a fish

source. FSANZ has assessed this aspect as part of the Safety Assessment (eport *Attachment+. "he Safety Assessment (eport confirms that #S itself is not allergenic but the yeaste&tract was allergic for se eral fish6allergic people tested. Beast allergenicity is notconsidered a food safety issue% nor is there a re-uirement of the $ode for yeast allergenlabelling within Standard 1. .7 H )andatory warning and ad isory statements anddeclarations. Se ere reactions to yeast ingestion is e&tremely rare% despite e&tensi e e&posureto common foods containing yeast. )ost indi iduals allergic to yeast appear able to toleratefoods containing yeast %0.

"!"!' /ene tec&nolo-# labellin- *rovisions

/i ision H abelling etc of food produced using gene technology in Standard 1.0. H Foodroduced using Gene "echnology% re-uires that processing aids and food additi es be labelled

where no el /NA andJor no el protein from the processing aid or food additi e remains present in the food to which it has been added.

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586 I// $/ a..r$//$. 9r / # // "/

"!0!1 Issues raised to t&e Initial Assessment +e*ort

ublic comment on the #nitial Assessment (eport was sought from 4

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;ueensland ,ealth noted an apparent inconsistency of the arguments supporting #S acting asa processing aid in the #nitial Assessment (eport% and the definition of a processing aid in

Standard 1.7.7. "he submitter underlined the rele ant wording to emphasis the point% whichis reproduced below.

#n the (egulatory roblem section of the #nitial Assessment (eport it was stated that:

9For #S in this Application to be considered a processing aid it needs to be performingits ma3or technological function during the processing or manufacture of the edible ice

products and no% or a minor% technological function in the final food.

2hile the definition of a processing aid is defined in Standard 1.7.7 as:

a substance listed in clauses 7 to 1 % where H

*a+ the substance is used in the processing of raw materials% foods oringredients% to fulfil a technological purpose relating to treatment or

processing% but does not perform a technological function in the final food!and

*b+ the substance is used in the course of manufacture of a food at the lowestle el necessary to achie e a function in the processing of that food%irrespecti e of any ma&imum permitted le el specified.

A number of submitters argued that #S is functioning more as a food additi e than a

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' Foods td states that currently stabilisers *food gums+ and emulsifiers are traditionallyused in ice cream manufacture to alter the ice cream te&ture and mouth6feel% and that these

products are considered food additi es. "he submission suggests that #S is performing asimilar function and should also be considered as a food additi e not as a processing aid.

"he AFG$ proposed a different iew% belie ing #S fulfils the re-uirements of a processingaid since it is performing its technological function during the freeIing process *manufactureof the ice products+ and has no technological function in the final food. "hat is% #S induces a

physical reaction during the freeIing process that modifies the ice crystal structure. "healtered ice crystal structure pro ides the changes in te&ture% fla our and colour retention. "heenhanced stability occurs because of the action of #S to alter the ice crystal structure during

processing and is not due to the presence of #S in the final product.

D /c // "

"he di ergent iews on this issue recei ed in submissions indicate that #S does not fit neatlyinto the usual processing aid categories. #t should also be pointed out that many food

additi es are also considered to be generally permitted processing aids. "his permission is pro ided by subclause 7*b+ of Standard 1.7.7. Food additi es listed in Schedule ofStandard 1.7.1 H Food Additi es can be considered to be generally permitted processing aidsif they meet the definitional re-uirements of Standard 1.7.7.

"here is a wide o erlap between food additi es and processing aids% and an importantcriterion used to decide into which group a substance best fits is how the compound is

performing its technological function! during processing and manufacture or in the final food.

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"hese two food additi e classes ha e -uite different mechanisms on the molecular le el tothat of #S . Stabilisers and emulsifiers do not ha e an effect on the ice crystal structure. "he

final effect may well be similar% that is amending final product te&ture% mouth6feel and melt properties% but the process is different.

For a substance to be considered a food additi e it needs to achie e a technological functionas listed within Schedule 0 of Standard 1.7.1. #S does not seem to fit neatly in any of thefood additi e functions listed in Schedule 0. ossible food additi e functions listed inSchedule 0 for #S are stabiliser or firming agent. Stabiliser includes binder% firming agent%water binding agent% foam stabiliser% and is defined as 9maintains the homogeneousdispersion of two or more immiscible substances in a food. Firming agent is defined as9contributes to firmness of food or interact with gelling agents to produce or strengthen a gel.#S does not conform to these Schedule 0 definitions or beha e in a manner of either astabiliser or firming agent.

#n this Application% #S is more correctly regarded as a processing aid rather than a foodadditi e.

0. .1.7 abelling issues

abelling re-uirements for the use of #S to produce ice cream and edible ices was raised asan issue in a number of submissions including that of the New Zealand Food SafetyAuthority. As stated in section 0. .1. abo e there was discussion about whether the proteinshould be considered a processing aid or food additi e for the proposed purpose of theApplication. "his decision has labelling implications as food additi es are re-uired to be

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Submissions from the /epartment of ,uman Ser ices Dictoria and aula Boung e&pressedthe iew that not labelling the presence of a substance produced from a genetically modified

source was misleading to consumers and did not allow consumers to ma5e choices onwhether to purchase product containing #S .

D /c // "

Standard 1. . 6 abelling of #ngredients% co ers the declaration of food additi es on labels in$lause and would apply if #S were to be regulated as a food additi e in Standard 1.7.1. #fa food additi e cannot be classified in one of the prescribed classes of food additi es% then itneeds to be listed by its prescribed name. #f it can be classified as one of the classes *such asfirming agent or stabiliser+ then it needs to be labelled with the name of the class followed bythe additi es specific name in brac5ets *or an #NS number% if applicable+.

"he >SA G(AS e&pert panel report for #S stated that the protein 9may be identified onlabels by the common or usual name of 9ice structuring protein. "he rele ant e&tract relatingto labelling is:

5&e ISP t#*e III *re*aration covered b# t&is /+AS evaluation ma# be identified on t&elabel of fro3en novelties sim*l# b# t&e common or usual name declared in t&edesi-nation of in-redients *ursuant to $1 CF+ 1 1!% (e!-!4 7ice structurin- *rotein8)!5&ere is no need for commercial *roducts to be labeled wit& t&e word 7fis&8 or an#ot&er desi-nation as a condition of safe use!

FSANZ has confirmed with the Applicant that commercial product produced using #S in the

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Along with the situation of chymosin mentioned in the #nitial Assessment (eport% thissubmitter belie es the #S situation is analogous to that of a number of enIyme processing

aids which are also deri ed from genetically modified micro6organisms% which are identicalto those from non6genetically modified sources and they also do not re-uire labelling underStandard 1.0. .

"wo submissions stated they belie ed there are consumer choice issues if labelling underStandard 1.0. is not re-uired% since this remo es consumers choice to ma5e decisions to not

purchase products that contain ingredients deri ed from genetically modified organisms.

D /c // "

Gene technology labelling re-uirements in Standard 1.0. do not apply to #S because #Sdoes not contain no el /NA nor is it a no el protein as defined in the Standard. "he #S ofthe Application is identical in amino acid se-uence to the counterpart #S found in nature*that is isolated from ocean pout+. "his is an analogous situation to that of chymosin *or other enIymes sourced from genetically modified micro6organisms+. "he chymosin sourced from

genetically modified sources is identical to that obtained from natural sources% and does notneed to be labelled under the re-uirements of Standard 1.0. . "he important point is whetherno el /NA or no el protein is in the final food. "he issue of whether #S is unchangedduring processing is irrele ant for labelling purposes. $heese is usually not heat processedafter the addition of chymosin% and the continued presence of undegraded chymosin has beenargued to be performing an additi e function as a fla ouring in ripened cheese.

"he situation for consumer choice concerning the identification of food that contains

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D /c // "

"his matter is addressed under section ? 6 #mpact Analysis.

0. .1. /rafting issues

"he AFG$ suggested that care would be needed to ensure correct legal drafting is pro ided toensure the Applicant achie es the permissions they are re-uesting% and G) permission orhigher than re-uested le els should be considered if the Application is accepted and there areno safety issues.

"he Applicant states that they wish to use #S during the manufacture of ice cream and edibleices% which includes products under item 7 of Schedule 1 of Standard 1.7.1. "he Applicantre-uested permission to include such products as froIen yoghurts and froIen fruit andJor

egetable 3uices and drin5s. "he submission states they belie e froIen yoghurt do notactually come under this item but would be considered a yoghurt% which has been froIen."he same situation e&ists for froIen fruit andJor egetable 3uices and drin5s.

D /c // "

#f #S is regulated as a processing aid% a drafting amendment is not re-uired for Schedule 1 of Standard 1.7.1. #f permission is pro ided for #S in Standard 1.7.7 H rocessing Aids%appro als can be pro ided to e&plicitly co er the product categories the Applicant hasre-uested. "he low le els of use proposed in this Application limit consideration of #S as afood or food ingredient and the safety assessment also considered the low le els of use

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#S functions as a processing aid as it is performing its technological purpose during themanufacturing step of ma5ing ice cream and edible ices% and does not perform a

technological function of a food additi e in the final products. #S 9binds to the de elopingice crystal structure and modifies it during formation. "he modified structure of the ice isresponsible for the stability of the products containing #S . "his is different in the case ofstabilisers and emulsifiers which are used in the traditional method of modifying mouth6feeland slowing product melt% where these chemicals act as food additi es since they ha e atechnological function in the final food consistent with Schedule 0 of Standard 1.7.1.

As a processing aid #S does not need to be labelled on final foods. #S also does not meetthe labelling re-uirements for substances that cause ad erse reactions to foods *Standard1. .7+ or foods deri ed from genetically modified sources *Standard 1.0. +.

#S is more consistently considered as a processing aid and therefore is most appropriatelyregulated within Standard 1.7.7. #S is not considered a food additi e since it does not

perform its technological function in the final food or meet one of the technological functionsof a food additi e in Schedule 0 of Standard 1.7.1. #t is not a stabiliser or a firming agent.

"here are two ris5 management options a ailable:

1. (egulate #S within "able to clause 1 H ermitted processing aids with miscellaneousfunctions of Standard 1.7.7 H rocessing Aids% gi ing specific functions of how it can

be used% for which products it could be used for and a detailed name of the protein% thatis #ce structuring protein "ype ### , $ 1 .

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8 I pac% A"a1(/ /

8 A99$c%$. Par% $/

"he affected parties to this Application include the following:

1. those sectors of the food industry wishing to mar5et the food products sub3ect to theApplication% specifically companies who wish to produce ice cream and edible ice

products!

. consumers! and

7. $ommonwealth% State% "erritory and New Zealand Go ernment agencies that enforcefood regulations.

82 I pac% a"a1(/ /

#n the course of de eloping food regulatory measures suitable for adoption in Australia and New Zealand% FSANZ is re-uired to consider the impact of all options on all sectors of thecommunity% including consumers% the food industry and go ernments. "he regulatory impactassessment identifies and e aluates% though is not limited to% the costs and benefits of the

proposed regulation% and its health% economic and social impacts.

"he following is an assessment by FSANZ of the costs and benefits of the two regulatoryoptions identified so far. "his is based on information supplied by the Applicant and

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Op% " 2

#ndustry: 'enefit to industry allowing the manufacture of new inno ati e and impro edice cream and edible ice products% especially in comparison to manufacturers inother countries where the technology is appro ed and has beencommercialised. Such possible new products could include low fat% low sugarand higher fruit products.

'enefit to importers and distributors of o erseas food products as the productrange is e&tended.

'enefit to industry groups in the supply chain of ice cream and edible ice products where new technology is a ailable to limit shelf life losses due tomelting of product.

'enefit to food retailers in an increased product range.

$onsumers: ossible benefit being able to purchase new inno ati e ice cream and edible ice products with impro ed sensory properties and impro ed shelf life of e&isting products *i.e. the products stay firmer longer and ta5e longer to melt+. Some possible new products with consumer benefits are low fat% low sugar and higher fruit products.

ossible cost may be paying a higher price for new premium inno ati e icecreams and edible ice products.

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$omments on% but not limited to% the following would be useful.

#s there technological 3ustification for the use of #S for the manufacture of ice cream

and edible ice productsL 2hat additional safety considerations would be associated with its proposed useL 2hat are the li5ely costs and benefits to food manufacturers% consumers and

go ernment if #S is appro edL 2ho are the affected parties relating to this ApplicationL

82 + r1. Tra.$ Or:a"

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"he proposed draft ariation to the $ode is consistent with the section 14 ob3ecti es ofthe FSANZ Act.

"o achie e what the Application see5s% namely permission to use #S as a processing

aid for the manufacture of ice cream and edible ices% there are no alternati es that aremore cost effecti e than a ariation to Standard 1.7.7.

ATTAC*MENTS

1! /raft ariations to the Australia New Zealand Food Standards Code$! Summary of public submissions'! Food technology report%! Safety assessment report"! /ietary modelling report

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A%%ach $"%

Dra9% ;ar a% "/ % %h$ Australia New Zealand Food Standards Code

T c $"c$ " :a

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A%%ach $"% 2

S ar( 9 p #1 c / # // "/

R ". O"$

S # %%$r Or:a" /a% " Na $1 Food "echnology Association of Dictoria /a id Gill

New Zealand Food Safety Authority $arole #n5ster

7 Australian Food and Grocery $ouncil "ony /owner ' Foods td )onica 2itsch0 /epartment ,uman Ser ice Dictoria Dictor /i aola

>nile er Australasia Culie Newlands? ;ueensland ,ealth Gary 'ielby

#ndi idual aula Boung

S # %%$r P / % " C $"%/Food "echnologyAssociation ofDictoria

"entati e support%with issues needingto be addressed.

"hey supported the Application% but they did ha e a numberof issues which they belie ed needed addressing. #S is not an acceptable abbre iation for the protein andshould not be used in any documentation% specifically not onlabels or ad ertising for any food products containing it.$onsumers would ha e no understanding of what #S is. "hey argue that #S has properties that ma5e it more li5e afood additi e than a processing aid.

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number of issues forconsideration

"hey ha e e&pressed the opinion that #S can beconsidered a food additi e% if it is ha ing a minortechnological function in the final food% which then re-uireslabelling in the ingredients list. "hey also suggest that consideration for the presence offish protein may re-uire labelling pro isions under Standard1. .7 *as #S is a fish protein sourced from a geneticallymodified yeast+.

Australian Food andGrocery $ouncil

Supports "he AFG$ supports the Application% sub3ect to a satisfactorysafety assessment."hey e&pect that #S will be considered safe due to:

#nternational appro als! established long6term human consumption! #S is a simple protein% and as such will be bro5en downand digested as any other protein! and other information supplied by the Applicant."hey also made a number of other comments. "hey belie e #S is a processing aid since it is performingits technological function during the freeIing process*manufacture+ and has no function in the final food. "heAFG$ states that #S induces a physical reaction during thefreeIing process that% together with the rate of the freeIing%modifies the ice crystal structure that is formed at sub6Ierotemperatures.

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manufacturers should not need to be passed onto consumers.)anufacturers should be able to produce new inno ati e

products while consumers should also benefit from impro ed-uality products. "here may be minor costs to go ernmentagencies if they need to perform analyses to chec5 forma&imum limits if such are imposed rather than G) .

' Foods td Support furtherconsideration butthey ha e a numberof issues andconcerns

As a manufacturer of ice cream and dairy products they ha ean interest in this Application. #n summary they supportfurther consideration of the Application% but they do ha e anumber of issues and concerns which they belie e need to beaddressed. $urrently arious stabilisers and emulsifiers are added toice cream to modify ice crystal growth and siIe which altersice cream te&ture and mouth6feel. "hey state these agents actsas processing aids but they also ha e a technological functionin the final products so act also as food additi es. "hey

belie e the same is the case with #S % that is it also has a foodadditi e function in the final food. "hey also ha e concerns about the allergenicity of #S % and belie e further ris5 assessment on the allergen aspects be performed. "hey belie e #S should be labelled on the final products.

/epartment of,uman Ser iceDictoria

/o not support theApplication

"hey belie e #S is a food additi e and can be considered astabiliser. "hey belie e the protein has an effect upon thete&ture of the final product% so has a technological function inthe final food. 'eing a food additi e it would be re-uired to

be labelled. #t is declared as an ingredient in the >SA. "hey belie e this Application would set a dangerous

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"hey ha e pro ided -uite a deal more informatione&plaining how ice cream *and edible ice products+ aremanufactured and how #S performs its technologicalfunction during manufacture. #S has no effect on thetemperature at which ice forms or the ice content% but it doesalter the siIe and shape of the crystals and so the final icestructure. "his information is to continue to 3ustify that they

belie e #S beha es as a processing aid for the proposed purpose not as a food additi e. "hey e&panded on the 3ustification for belie ing that #Sdoes not need to be labelled under Standard 1.0. . "heyreiterated that they belie e #S is not a no el protein% since itis identical to a fish protein which has a history of safe use.Also they state the situation is analogous to that for enIymes*including chymosin% as stated in the #nitial Assessment+which ha e been deri ed from genetically modified organismswhich do not re-uire labelling under Standard 1.0. .

;ueensland ,ealth No position at thisstage but made somecomments

"hey stated they neither accept nor re3ect the Application atthis stage but will re iew once the safety assessment*including allergenicity aspects+ has been performed.

,owe er they did point out an inconsistency in the #nitialAssessment (eport *#A(+ 3ustification of #S acting as a processing aid and the definition of a processing aid in the$ode."he #A( stated *underlined in the submission to highlight thedifferences+:9For #S in this Application to be considered a processing aidit needs to be performing its ma3or technological functionduring the processing or manufacture of the edible ice

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A%%ach $"% 3

F . T$ch" 1 :( R$p r%

G$"$ra1 I"%r . c% " 9 r Ic$ S%r c% r ": Pr %$ "/

$ells of li ing organisms are usually irre ersibly damaged during freeIing causing cell death.FreeIing depri es cells of their a-ueous medium which they re-uire for functioning% causesion and solute concentration in the plasma% causes denaturation of biomolecules and can

rupture cell membranes *,arding et al! % 1888+. ,owe er a number of arious organismsincluding fish% plants% insects% fungi and bacteria ha e been identified that are able to sur i eat temperatures below freeIing *'arnett% 441+. Such di erse organisms ha e been found tocontain molecules *essentially proteins and peptides+ which assist sur i al by depressing thefreeIing point of cell li-uids. < er thirty years of research has been performed on these

proteins. Such proteins were first identified in 18 8% in the blood of fish li ing in areas wherethe sea froIe */e Dries and 2ohlschlag% 18 8+.

"hese proteins ha e been gi en arious names such as antifreeIe proteins% ice growthmodifiers% thermal hysteresis proteins and now more recently ice structuring proteins *$lar5eet al! % 44 +. "he term ice structuring proteins has been proposed because regardless of theirsource and structure all the proteins bind to and influence the growth of ice crystals.

"he term thermal hysteresis is defined as *,arding et al % 1888+:

the difference between

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"he summary of the agreed understanding is that the ice structuring protein 9binds to thede eloping ice crystal in one particular a&is% so limiting growth in this direction. Also it is

belie ed that ice structuring protein adsorbs preferentially onto a specific face of thede eloping ice crystal. "here is a ariety of -uite detailed analyses of possible mechanismsfor binding. "hese analyses detail crystal structure geometries and the arious proteins Q6raycrystal structures of classes of ice structuring proteins but for the purposes of the Applicationit is sufficient to 5now that ice structuring proteins accumulate *if not strictly chemically9bind+ to specific faces of the ice crystal and so alters the growth patterns and growth rates of the ice structures. "his alteration also changes the physical properties of the ice productsformed by their use in commercial ice products *discussed below+. #t has been postulated thatthe adsorption of the proteins on the ice crystal structure is due to fa ourable intermolecularsteric interactions and an der 2aals forces. #t is also belie ed that both hydrophobic andhydrophilic interactions are in ol ed.

Sp$c 9 c #ac :r ". " %h$ c$ /%r c% r ": pr %$ " 9 %h$ App1 ca% "

"he arious ice structuring proteins which ha e been identified from a ariety of different

organisms ha e been classified into different groups which ha e similar protein structuresand properties. For ice structuring proteins isolated from fish arieties the groups ha e beentermed type #% ##% ### and #D *$re elet al! % 44 +. "he ice structuring protein of thisApplication is categorised as a type ### protein. Fish type ### ice structuring proteins ha e

been found in the following fish: ocean pout% eelpout and wolffish.

"he ice structuring protein of this Application was originally isolated and is found naturallyin ocean pout * 2acro3oarces americanus +% which is a cold water fish found off the northeast

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)ore detailed discussion about the molecular biological and safety aspects% including thegenetic stability of the modified yeast% are contained in the Safety Assessment (eport

*A%%ach $"% 4+.

"he commercial production of #S from the modified yeast occurs using standard industrialscale batch fermentations% with subse-uent isolation using microfiltration% concentration and

pac5aging steps. "his is ery similar to production processes for commercial enIymes usedfor food manufacture. "he commercial #S preparation is a mi&ture of #S % glycosylated #S*#S bound to the sugar mannose+% proteins and peptides from the yeast and sugars% acids andsalts commonly found in food. "he #S preparation is standardised and stabilised in citricacid buffer.

T$ch" 1 : ca1 /% 9 ca% " 9 r ISP

As mentioned in an abo e section ice structuring proteins affect the growth and structure ofice crystals by directly accumulating or adsorbing *if not strictly chemically 9binding+ to thegrowing ice crystals and inhibiting the crystal growth *particularly in one direction or a&is+

resulting in modification of the resulting ice crystal siIe and structure and hence its physical properties. For food products based on ice% addition of #S also has important impacts on thesensory properties of the resultant ice products. Such altered sensory properties includeresultant hardness *and how long before the ice product melts+% creaminess and alterations tofla our deli ery. "hese aspects were postulated in some of the recent references concerningice structuring proteins *specifically Griffith and @wart% 1880+.

Some of the suggested ad antages of using #S during the manufacture of ice cream and

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$ommercial manufacture of ice cream or edible ices occurs in a standard freeIer where cold icecream or water ice mi& enters and is cooled on the cold walls of the freeIer. "he ice% which

forms on the walls% is scraped off bac5 into the mi&ture. Nearly all the ice crystals present in thefinal products are formed in the freeIer stage. "he ice crystalsJwater mi& continues through thefreeIer stage where the ice crystals formed increase in siIe. #t is stated that typical manufactureof ice cream and edible ices has the product mi& entering the freeIer at 0P$ and e&truded atappro&imately H P$ where appro&imately 4K of the final ice structure has been formed.$older e&truder temperature increases the percentage of ice formed.

/uring the freeIer stage the addition of #S alters the shape and siIe of the ice crystals! withcrystals produced with the addition of #S being rod shaped rather than the usual roundshape. "he resultant smaller rod shaped ice crystals produce a product from the e&truder thatis firmer and has higher iscosity *see pictures below in Figure 1 pro ided by the Applicant intheir submission to the #nitial Assessment (eport+.

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"he situation with using #S is different to the technology% which is traditionally used to alterthe physical properties of ice cream and edible ice products including te&ture% mouth6feel and

melt resistance. "he traditional method uses food additi es called stabilisers *food gums+ andemulsifiers to alter the properties. Stabilisers alter the iscosity of the ice cream matri&%which modify the gel networ5 at the interface between the ice structure and the water matri&."his increased iscosity slows down the diffusion during melting so slowing down meltingeffects. @mulsifiers impro e the miscibility of two different phases! water and fat in the icecream mi&ture. @mulsifiers also impro e stability of air bubbles in mi&tures where air isadded to ice cream products to impro e their properties.

Sp$c 9 ca% " 9 ISP

"he Applicant states that there is no international standard for #S . "hat is there is no $ode&standard% and C@$FA *Coint FA

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Sp$c 9 ca% " 9 r c$ /%r c% r ": pr %$ " %(p$ III *PLC 2 pr$para% "8

#ce structuring protein type ### , $ 1 preparation is a protein e&creted from thefermentation of a genetically modified yeast * Sacc&arom#ces cerevisiae + to which a syntheticgene encoding for the protein has been inserted into the yeasts genome.

Assay Not less than 0 gJ acti e ice structuring protein type ###, $ 1

p, 7.4 J64.0Ash Not more than K

Appearance ight brown a-ueous preparation,ea y metals Not more than mgJ)icrobial limits

"otal microbial count O7444 per g$oliforms O14 per gBeast and mould count O144 per g

9isteria s*! Absent in 0 gSalmonella s*! Absent in 0 g

:acillus Cereus O144 per g

C "c1 / "

"he Application to use #S as a processing aid during the manufacture of ice cream andedible ices is technologically 3ustified.

R$9$r$"c$/

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A%%ach $"% 4

Sa9$%( a//$// $"% r$p r%

APPLICATION A544 H ICE STRUCTURING PROTEIN AS A PROCESSING AID INICE CREAM AND EDIBLE ICES

SUMMAR> AND CONCLUSIONS

Bac :r ".#ce Structuring rotein type ### , $ 1 *#S +% deri ed from a northern hemisphere fishspecies% has been assessed in terms of safety for human consumption. Naturally occurring icestructuring proteins can bind to and influence the growth and structure of ice crystals%resulting in a modified ice structure. 2hen used in the manufacture of certain froIen food

products% these properties affect the physical and sensory properties of the foods% as well asimpro e temperature stability. #n this application% permission is sought to use #S type ###

, $ 1 as a processing aid in the manufacture of dessert products such as ice cream andwater ices.

As natural fish sources are limited% the Applicant has de eloped a method of producingcommercial -uantities of #S by fermentation of ba5ers yeast that has been geneticallymodified *G)+ to manufacture and secrete the fish #S . "he #S preparation is a mi&ture offunctionally acti e #S % inacti e mannose6con3ugated #S % proteins and peptides fromcommon ba5ers yeast% and sugars% acids and salts commonly found in food.

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S5in pric5 testing with #S did not produce any positi e reactions to the protein% althoughfour reactions to yeast proteins were obser ed and confirmed by in vitro tests. A

confirmatory s5in pric5 test with a highly purified #S *yeast protein content O1K+ wasnegati e. "he conclusion from these in estigations was that #S is not li5ely to be allergenicin humans.

#n studies using human olunteers% ingestion of #S preparation for eight wee5s at a highdaily dose did not result in specific antibody formation% indicating that #S is not li5ely to beany more immunogenic than the ma3ority of dietary proteins.

Additional biochemical analyses simulating gastric fluid digestion with pepsin in an in vitro test system showed that both #S and its glycocon3ugated form would be readily degraded inthe human digesti e system. #n addition% amino acid se-uence analysis showed asusceptibility to proteolytic brea5down by intestinal enIymes such as trypsin. "hese resultsindicate that #S is therefore unli5ely to be absorbed intact or accumulate in the body. 'ased on a thorough assessment of allergic potential% and the results of the analytical% animal%

human% andin vitro data presented in this application% #S preparation is not to&ic and isunli5ely to e o5e an allergic reaction in fish6sensitised indi iduals% or to sensitise potentiallysusceptible indi iduals in the wider population.

C "c1 / "

No potential public health and safety concerns ha e been identified in the assessment of #S .

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28 *ISTOR> OF USE

"he Applicant states that hundreds of 5ilograms of #S would be re-uired each year togenerate commercial -uantities of froIen dessert products. nited States Food and /rugAdministration *>S F/A+ system. #n 188 % the >S @n ironmental rotection Agency *@ A+e aluated the ris5 associated with industrial use of Sacc&arom#ces cerevisiae % including G)strains% and concluded that human health and en ironmental release ris5s associated with thisorganism are low% and that it poses no significant health haIard.

282 D " r r:a" /

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"hus consumption of a 44g portion of ocean pout would result in an inta5e of between 1 4mg and 4 mg of #S type ### from the diet. Fletcher et al . *18 0+ reported that ice

structuring proteins are present in fish plasma all year round% and therefore consumption ofocean pout would always be associated with consumption of #S type ###.

38 DESCRIPTION OF T*E GENETIC MODIFICATION

38 M$%h . /$. " %h$ :$"$% c . 9 ca% "

"he gene e&pression cassette encoding #S type ### , $ 1 *deri ed from ocean pout+ wasintroduced into ba5ers yeast using osmotic shoc5% which increases the permeability of theyeast cell membrane allowing the upta5e of e&ogenous /NA. "he gene cassette is then ableto automatically integrate into the yeast chromosomal /NA% at the ribosomal /NA *r/NA+locus% as a stable% multi6copy insert.

Strain descri*tion

roducing strain: $@N E77 containing multi6copy integration fragment of plasmid p>(7887 integrated at the r/NA locus. *$@N E77 R Sacc&arom#ces cerevisiae 2A5a 2A9$

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383 M 1$c 1ar charac%$r /a% " 9 %h$ ($a/%

Insert and co*# number

Southern blot analysis was used to establish the site of integration of the inserted genecassette and the number of copies. "he presence of multiple copies shows that the integrationhas been targeted towards the ribosomal /NA locus as intended.

(8778 has occurred at ther/NA locus in the yeast genome.

384 S%a# 1 %( 9 %h$ :$"$% c cha":$

Genetic stability of the #S 6modified strain of S! cerevisiae was measured after more than ?4generations of growth under non6selecti e conditions. lating cells on selecti e and non6selecti e media re ealed the same amount of iable cells. #nducti e growth *after ?4

generations+ showed identical e&pression le els of #S when tested in li-uid culture.olymerase chain reaction * $(+ analysis on whole yeast cells *chromosomal /NA astemplate+ demonstrated that the #S gene was present. #n addition% Southern blot analysisshowed that the strain after ?4 generations was identical to the initial modified strain withrespect to the integration site.

"hese results demonstrate that the genetic modification in the engineered yeast strain isstable.

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Ta#1$ D / ": "9 r a% " 9 r %$/% a". c "%r 1 :r p/ " %h$ 3- $$ ra% /% .(

Gr p ISP %(p$ III *PLC 2 T %a1 ISP T %a1 S 1 ./2ater control 4 mgJ5gJday 4 mgJ5gJday 4 mgJ5gJday$itric acid control 4 mgJ5gJday 4 mgJ5gJday 144 mgJ5gJday

ow dose 0 mgJ5gJday 144 mgJ5gJday 44 mgJ5gJday#ntermediate dose 84 mgJ5gJday 4 mgJ5gJday 444 mgJ5gJday,igh dose 0 4 mgJ5gJday 8 4 mgJ5gJday 444 mgJ5gJday

arameters measured in the study included clinical obser ations% food consumption% neuro6 beha ioural testing% opthalmoscopic e&amination% clinical pathology *haematology% clinicalchemistry% urinalysis% bone marrow smears+% gross necropsy% selected organ weights andhistopathology of specified organsJtissues.

Summar# of e *erimental observations

$linical signs: Animals were obser ed daily for signs of ill health or o ert to&icity.Additional obser ations were conducted daily during 2ee5 1immediately post dosing% and 74 minutes% 1% % and hours afterdosing. ost dosing obser ations were made once wee5ly after 2ee51.

hysical e&amination: erformed at wee5ly inter als

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:acterial +everse 2utation Assa#

"he bacterial re erse mutation assay was performed using Salmonella t#*&imurium histidine6re-uiring strains "A1070% "A107?% "A8 % "A144% and "A14 and was compliant with

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/ene 2utation Assa# usin- 2ouse 9#m*&oma 9"1@. Cells

Gene mutation was assessed using the tmidine ,inase (t,) locus in mouse lymphoma01? B cells and was compliant with

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"he study in ol ed the participation of si&ty6nine healthy men and women who met particular age and health criteria determined at the commencement of the study. "he

participants recei ed a single ser ing of either control food or test protein food at wee51% andthe opposite product at wee5 *cross6o er+. "he control and test products were designed to be as similar as possible in composition.

Clinical monitorin- of subBects

"he safety and acceptability of the test material were assessed by monitoring treatment6emergent ad erse e&periences in the study participants% at each clinic isit *2ee5s 1 and +.At the screening isit *2ee5 4+ and 6hours following study product ingestion at eachtreatment clinic isit *2ee5s 1 and +% clinical laboratory testing% including serum chemistryand haematology profiles% were performed. Dital signs were measured at the screening isit*2ee5 4+ and prior to and 6hours following study product ingestion at each treatment clinic

isit *2ee5s 1 and +. At the screening isit *2ee5 4+% a urine sample was collected forroutine testing *all sub3ects+ and for a pregnancy test *all females of childbearing potential+.At the screening isit *2ee5 4+ and at the end of the study *2ee5 +% a brief physicale&amination was conducted.

+esults and conclusion

"here were no significant differences in the test product containing #S and the control product in terms of effects on serum chemistry% haematology% ital signs% or occurrence ofad erse e ents. "hese results indicate that a single ingestion of yeast6deri ed #S in fooddoes not elicit ad erse reactions in otherwise healthy adults.

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"he applicants assessment of the potential allergenicity of #S has considered two issues: *i+whether the protein is li5ely to sensitiIe potentially susceptible indi iduals and thereby

increase the li5elihood of a reaction on subse-uent e&posure to that protein% and *ii+ whetherthe protein is li5ely to pro o5e a reaction in indi iduals allergic to the source from which the protein originated *or to structurally related proteins+. "his approach is consistent with recentinternational consensus documents% including the recommendations of a recent FA

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Amino acid se-uence analysis can identify regions in the linear se-uence of a protein thatresembles the se-uence of 5nown allergens. "he absence of any similarity suggests that a

protein does not possess any possible se-uence epitopes resembling those present in 5nownallergens. Se-uence analysis can also indicate whether the protein shares any structuralsimilarity with classes of proteins containing 5nown allergens and thus pro ide guidance forsubse-uent serum screening.

Se eral algorithms ha e been proposed for this purpose% but the most fre-uently used areFAS"A and ' AS" *'asic ocal Alignment Search "ool+% from which computer programs of the same name ha e been generated. 'oth methods rely on assessing the probability that analignment between a -uery se-uence *the un5nown protein+ and a se-uence in the databaseoccurs by chance. "he FAS"A program automatically searches for and eliminates regions oflow comple&ity% for e&ample multiple repeats of one or two amino acids% which wouldotherwise result in apparently significant similarity% but without necessarily ha ing any

biological significance. >sing ' AS"% as for the FAS"A program% low comple&ity regions%which would be e&pected to gi e ery high alignment scores without biological significance%are screened out.

Se-uence analysis of #S type ### was performed in line with the suggested procedures *FAnited Nations% (ome.

FA< * 441+. @ aluation of allergenicity of genetically modified foods. (eport of a Coint FA

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FA

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A%%ach $"% 5

D $%ar( $ p / r$ a//$// $"% r$p r%An application was recei ed by FSANZ from >nile er Australia imited re-uestingamendment of Standard 1.7.7 H rocessing Aids of the Australia New Zealand FoodStandards Code *the $ode+ to include the use of #ce Structuring rotein "ype ### , $ 1*#S + as a processing aid for the preparation of ice cream and edible ices. @dible ices includefroIen yoghurts and froIen fruit andJor egetable 3uices and drin5s.

A dietary e&posure assessment was deemed necessary in order to determine the estimateddietary e&posure to #S for the Australian and New Zealand populations if #S were added toice creams and edible ice products.

S ar(

A dietary e&posure assessment was underta5en to estimate dietary e&posure to #S for the

Australian and New Zealand populations. "he population sub6groups e&amined were thewhole population * years and abo e for Australia! 10 years and abo e for New Zealand+%toddlers * 6 years for Australia+% primary school aged children *061 years for Australia+%and teenagers *17618 years for Australia! 10618 years for New Zealand+. Food consumptiondata based on the 1880 National Nutrition Sur ey *NNS+ and 188? New Zealand NNS wereused to estimate #S dietary e&posure.

The estimated mean dietary exposures for consumers of ISP for Australia

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The 95 th percentile dietary exposures for consumers of ISP for NewZealand were estimated as:

mgJday for the whole population aged 10 years and abo e! and 7 mgJday for teenagers aged 10618 years.

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S F/A *Food and /rug Administration+ has deemed this #S as generally recognised assafe *G(AS+. $ommercial ice creams and edible ices incorporating #S ha e been sold in

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"he Applicant pro ided an Acceptable /aily #nta5e for #S of 0. mg #S J5g bwJday.,owe er% neither the Coint FA

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* $r$ %h$ $/% a%$. . $%ar( $ p / r$/ ca1c 1a%$.J

"he /#A)

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all the foods within the group contain #S at the proposed le els specified in "able 1.>nless otherwise stated% the ma&imum concentration of #S in each food category has

been used! consumption of foods as recorded in the NNSs represent current food consumption patterns!

consumers always select the products containing #S ! consumers do not alter their food consumption habits to substitute non6#S containing

products with #S containing products! consumers do not increase their consumption of foodsJfood groups upon foodsJfood

groups containing #S becoming a ailable! all #S present in food is absorbed by the body! naturally occurring sources of #S ha e not been included in the dietary e&posure

assessment! where a food was not included in the e&posure assessment% it was assumed to contain

a Iero concentration of #S ! and where a food has a specified #S concentration% this concentration is carried o er to

mi&ed foods where the food has been used as an ingredient e.g. ice cream used in thic5sha5es.

"hese assumptions are li5ely to lead to a conser ati e estimate for #S dietary e&posure.

L %a% "/ 9 %h$ . $%ar( .$11 ":

A limitation of estimating dietary e&posure o er a period of time associated with the dietary

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R$/ 1%/

E/% a%$. . $%ar( $ p / r$/ % ISP

"he estimated mean and 80 th percentile dietary e&posures for consumers of #S in Australiaand New Zealand are shown in Figure 1 *mgJ5g bwJday+ and Figure *mgJday+.

@stimated #S dietary e&posures are presented for consumers of #S only and not for allrespondents *e ery person in the population group+. For details on the number of respondentsand consumers in each population group assessed% see "able A1.1 in Appendi& 1.

The estimated mean dietary exposures for consumers of ISP in Australiawere:

1 mgJday *4. mgJ5g bwJday+ for the whole population aged years and abo e! mgJday *4.0 mgJ5g bwJday+ for toddlers aged 6 years! 17 mgJday *4. mgJ5g bwJday+ for primary school children aged 061 years! and

1? mgJday *4.7 mgJ5g bwJday+ for teenagers aged 17618 years.

The estimated mean dietary exposures for consumers of ISP in NewZealand were:

14 mgJday *4.1 mgJ5g bwJday+ for the whole population aged 10 years and abo e! and 10 mgJday *4. mgJ5g bwJday+ for teenagers aged 10618 years.

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F : r$ 8 E/% a%$. $a" a". 75 %h p$rc$"% 1$ . $%ar( $ p / r$/ 9 r c "/ $r/ 9 ISP ! : : # .a(' 9 r ;ar / A /%ra1 a" a". N$,$a1a". p p 1a% " :r p/8

4.

4.0 4.

4.7

4.1 4.

4.?

1.7 1.

4.8

4.

4.G

4.4

4.

4.

4.G

4.F

1.4

1.

1.

years and abo e 6 years 061 years 17618 years 10 years and abo e 10618 years

A0/%ra1&a N$2 ,$a1a". P p01a%& " /0#-:r 0p/

E / %

& a % $ .

I S P . & $ % a r ( $ 3 p - / 0 r $

! : D

B :

# 2

D . a ( '

)ean

80th :ercentile

?

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F : r$ 28 E/% a%$. $a" a". 75 %h p$rc$"% 1$ . $%ar( $ p / r$/ 9 r c "/ $r/ 9 ISP ! : .a(' 9 r ;ar / A /%ra1 a" a". N$ ,$a1a".p p 1a% " :r p/8

1 F

17 1?

14

10

77

7

7

8

G

7F

4

14

4

74

4

04

G4

years and abo e 6 years 061 years 17618 years 10 years and abo e 10618 years

A0/%ra1&a N$2 ,$a1a". P p01a%& " /0#-:r 0p

E / % & a % $ .

I S P . & $ % a r ( $ 3 p - / 0 r $

! : D

. a (

'

)ean 80th :ercentile

?8

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REFERENCES

#nstitute of )edicine% National Academy of Sciences% 444% ietar# +eference Inta,es for Eitamin C4Eitamin 64 Selenium4 and Carotenoids % National Academy ress% 2ashington% /$.

National ,ealth and )edical (esearch $ouncil% 441% National ?ealt& and 2edical +esearc&Council ebsite ( www!&ealt&!-ov!auGn&mrcG*ublicationsGdiet )

(utishauser #. 444. /ettin- it ri-&t>< &ow to use t&e data from t&e 1 " National Nutrition Surve# .$ommonwealth of Australia: $anberra

2orld ,ealth

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