Reducing the levels of cyanogenic glucosides in cassava Hazel Fuchs, 2008 Cassava kills!

! It is well known that plants such as aconite, hemlock and belladonna are hi hl! poisonous, but man! people are not aware that there are certain t!pes o" staple "ood which can also be classi"ied as to#ic$ %ome "oods which have been eaten "or centuries b! humans can even cause serious poisonin $ Cassava tubers are a strikin e#ample o" this dilemma$ &he! have been eaten ever since mankind learnt to cook$ 'eople are inclined to believe that because humans have been usin this crop since the dawn o" time it must be sa"e$ (ut the time has come to put an end to this misconception$ )an! edible plants have been "ound to contain various poisonous substances which a""ect human metabolism, blockin important

biochemical pathwa!s* one o" these hi hl! potent poisons is linamarin, a c!anide derivative which is hi hl! concentrated in cassava +Manihot esculenta,$ Cassava, also known as manioc, is an important staple "ood in -"rica, %outh -merica and %outh .ast -sia, contibrutin to up to 80/ o" the people0s diet, especiall! in (razil$ It is a known cause o" chronic c!anide poisonin and in a "ew cases is "atal$ &he 1uestion is how it can be modi"ied to make it less poisonous and the answer is to sieze the problem b! the roots and manipulate the plant0s enes to make it sa"e "or consumption$ &he "irst aspect which should be considered when discussin the to#icit! o" M.esculenta is which compounds make it so poisonous$ It has been

Hazel Fuchs

shown that the main compounds that cause the to#ic e""ect are linamarin and lotaustralin +%in h et al$, 2384,, which are c!ano enic lucosides$ C!ano enic lucosides are usuall! 5* lucose 6oined b! a *linka e to an acetone c!anoh!drin derivative +78r ensen et al$, 2009,$ &he! are s!nthesized b! the plant to protect it "rom predation, as h!dro en c!anide, which is released "rom these compounds when the! are di ested, is poisonous to virtuall! all known eukar!ote li"e "orms +%in h et al$, 2384,$ M. esculenta produces the c!ano enic lucosides in a ver! speci"ic wa!$ &he "irst step is to convert :*valine into +;,*2*meth!lpropanal*o#ime$ &his is catal!sed b! two similar c!tochromes +'<90s, which are encoded b! the enes C='>352?52$ +-ndersen et al$, 2000,$ &here are two o" these enes because M.esculenta is an allopol!ploid$ In the ne#t step +;,* 2*meth!lpropanal*o#ime reacts to acetone c!anoh!drin and in the "inal step a lucose molecule is bound to the acetone derivative and linamarin is "ormed$ &he e#pression o" the enes C='>352 and C='>352 varies reatl! in the appro#$ 9000, at least phenot!picall! di""erent t!pes o" cassava, makin their c!ano enic potential +C@', ran e "rom 20*900 ppm$ &he plants can be rouped accordin to their c!ano enic potential +C@',, which is e1uivalent to their c!anide content$ &here are low*C@' plants +less than 90 ppm o" c!ano enic e1ivalents, and hi h*C@' plants +more than 200 ppm,$ Aenerall! the low*C@' tubers, which row durin a !ear with normal rain"all levels, can be cooked as the! are, whereas the hi h* C@' roots Hazel Fuchs rown durin a drou ht need to be processed prior to 2

consumption$ -ll o" the rou hl! 29 di""erent t!pes o" c!ano enic lucoside consist o" a lucose molecule which is *linked to an acetone c!anoh!drin$ &he onl! di""erence between them lies in the residal roup which is bound to the end o" the acetone c!anoh!drin$ In the case o" linamarin it is a h!dro en atom, and "or e#ample, in lotaustralin a meth!l +**CHB, roup$ -ll c!ano enic lucosides are metabolized in similar wa!s b! predatorsC &he cells o" the tubers which contain, "or e#ample, linamarin also contain a * lucosidase which is released simultaneousl! with the c!ano enic lucoside$ &he enz!me di ests the substrate b! releasin c!anide "rom the c!ano enic roup which is then protonated to h!dro en c!anide

+HC@,$ &his process continues until the * lucosidase is denatured b! the low pH due to the h!drochloric acid in the stomach$ &he reason wh! HC@ is so poisonous is that it binds to the Fe 2D in hemo lobin "ormin c!anohemo lobin +Cereda E )attos, 2334,$ -s the bindin a""init! o" c!anide is much hi her than the e1uivalent bindin a""init! o" o#! en, the respirator! c!cle is impeded$ Hi h levels o" h!dro en c!anide in the bloodstream lead to su""ocation o" the cells and result in acute poisonin with s!mptoms such as sickness, diarrhoea, stomach ache, headache and, in some cases, even death$ &his makes HC@ and also the c!ano enic lucosides so dan erous$ :ow levels o" HC@ can be deto#i"ied slowl! b! a mechanism which has been e#amined in pi s +Fke, 2343,, but usuall! the er!throc!tes containin c!anohemo lobin 6ust circulate uselessl! in the blood stream until the! a e enou h to be broken down$ &his lowers the o#! en transport capacit! o" the blood and i" it is too low this can

Hazel Fuchs

result in chronic poisonin causin serious illnesses such as Gonzo +a "orm o" paral!sis, or &ropical -ta#ic @europath! +&-@, +Cardoso et al$,200B,$ )an! e""orts have been made to reduce the to#icit! o" cassava$ First o" all, there are traditional methods o" preparin the tubers "or cookin $ %un*dr!in and heap "ermentation o" the tubers, used "or e#ample in )ozambi1ue and parts o" western -"rica, are the most e""ective$ &hese reduce the amount o" linamarin in cassava to appro#$ 29/ and 29/, respectivel!$ Aratin the roots be"ore their preparation is use"ul, because the cells are dama ed more and then the linamarin reacts more easil! with the enz!me linamarase$ Fther methods have been tried such asC boilin , steamin , "r!in and bakin o" the roots$ &he water*based

preparation is less e""ective as the linamarase denatures at 200HC and is there"ore deactivated$ %teamin , "r!in and bakin cassava have ver! little e""ect on the c!anide content, as the linamarase is incapable o" di estin linamarin in the neutral to sli htl! acidic conditions that these methods create$ In most re ions o" -"rica cassava is prepared b! sun* dr!in or heap "ermentation, but in a plant with a rather hi h C@' even the preparation b! heap "ermentation leaves a residue amount o" appro#$ >9 ppm o" linamarin per k o" cassava +Cardoso, 200B,$ &his still is 490/ above the IHF sa"e level "or c!ano enic lucosides +Cardoso, 200B, in manioc "lour$ Consumption o" these products would lead to serious health problems$ Fbviousl! these methods need to be improved reatl! to prevent a diet based on cassava causin lon *term poisonin $ &he second possibilit! o" ensurin that onl! sa"e "lour is eaten is to

Hazel Fuchs

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provide the people who prepare cassava tubers with preliminar! tests indicatin c!anide levels in the root mash be"ore it is dried to make "lour$ &hese tests show whether or not the preparation o" the "ood is ade1uate "or consumption$ F" course these tests could onl! be made available to all i" lar e sums o" mone! were provided b! sponsors, and even then man! people mi ht i nore the tests i" the! had onl! a limited amount o" crop due to drou ht$ &he third and most promisin possibilit! to solve this problem is one which has onl! recentl! been e#amined b! %iritun a and %a!reJ to eneticall! en ineer cassava plants to produce less c!ano enic lucosides b! deactivatin selectivel! switchin o"" the certain enes$ &he team succeeded in

enes C='>352?52 which encode the enes$

c!tochromes '<90s b! antisense e#pression o" the respective

&hese enz!mes catal!se the "irst step o" the linamarin s!nthesis$ %iritun a and %a!re used promotors which are onl! "ound in either the roots o" the plants +cab2, or in the leaves +patatin,$ &his method provided selective reduction in leaves or tubers o" up to 3</ o" c!ano enic lucoside

content +%iritun a E %a!re, 200B,$ 'revious studies show that linamarin can be produced in the roots o" M.esculenta as well as in the leaves and that the plant transports it "rom lea" to root$ &here"ore stoppin the ene e#pression in the roots alone is not su""icient +%iritun a and %a!re 200<,$ %adl! the plants which s!nthesize less linamarin are unable to row in soil with a reduced ammonia +@HB, content, su the c!ano enic estin that the! re1uire L@-

lucosides in their nitro en c!cle$ Ksin

inter"erence technolo ! 78r ensen et al$ +2009, succeeded in reducin

Hazel Fuchs

linamarin levels in leaves b! more than 33$3/ and b! up to 32/ in tubers$ Aenetic en ineerin has been the most promisin method so "ar because once trans enic plants have been manipulated to produce e#tremel! low levels o" linamarin, the! will provide a reater 1uantit! o" sa"e "ood than preliminar! tests or a "oolproo" method o" cassava preparation can ensure, even i" the! demand more nutrients than the wild t!pe$ -s cassava tubers contain these potentiall! poisonous compounds, it is vital that their use as a "ood source be considered$ F" course, in man! &hird Iorld countries where "amine is a common problem, convincin the natives to starve instead o" eatin potentiall! poisonous roots is not an option$ (ut it is crucial that the cassava*eatin nations become more aware o" the conse1uences that wron preparation o" their "ood can

entail$ Fn the other hand these risks can easil! be reduced b! rowin trans enic manioc plants$ &hus I appeal "or more research into the metabolism and bios!nthesis o" c!ano enic lucosides J it could save lives! References: -ndersen, )$5$ et al$ +2000,C C!tochromes '*<90 "rom cassava +Manihot esculenta Crantz, catal!sin the "irst steps in the bios!nthesis o" the c!ano enic lucosides linamarin and lotaustralin$ 7 (iol Chem 2>9, p$ 2344J23>9$ Cardoso, -$ .t al +200B,C 'rocessin o" cassava roots to remove c!ano ens$ 7ournal o" Food Composition and -nal!sis 28, p$ <92J<40$ Cereda, )$ '$ and Mattos, M.C.Y. (1996): Lina arin ! "he to#ic Hazel Fuchs 4

co $ound of cassava. 7ournal o" Menomous -nimals and &o#ins, vol$ 2 n$2$ Food %tandards -ustralia @ew ;ealand +F%-@;, +200<, C C!ano enic l!cosides in cassava and bamboo shoots* - human health risk assesment, &echnical Leport %eries @o$ 28$ 78r ensen, G$ et al$ +2009,C Cassava 'lants with a 5epleted C!ano enic Alucoside Content in :eaves and &ubers$ 5istribution o" C!ano enic Alucosides, &heir %ite o" %!nthesis and &ransport, and (locka e o" the (ios!nthesis b! L@- Inter"erence &echnolo !$ 'lant 'h!siolo !, %eptember 2009, Mol$ 2B3, p$ B4BJB><$ Fke, F$:$+2343,C &he role o" h!droc!anic acid in nutrition$ Iorld Lev$ @utr$ 5ietetics$, vol$ 22, p$ 2>0*38$ %in h, H$ ($ et al$ +2384,C 5etection and determination o" c!anide* review$ International 7ournal o" .nvironmental -nal!sis o" Chemicals 24, p$ 229J2B4$ %iritun a, 5$ N %a!re, L$ +200<,C .n ineerin c!ano en s!nthesis and turnover in cassava +Manihot esculenta,$ 'lant )olecular (iolo ! 94, p$ 442J443$ Iilson, I$)$ +200B, C Cassava +Manihot esculenta Crantz,, C!ano enic 'otential, and 'redation in @orthwestern -mazoniaC &he &ukanoan 'erspective$ Human .colo !, Mol$ B2, @o$ B, %eptember 200B + CH 200B,

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