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Chloroplast‐derived enzyme cocktails hydrolyse lignocellulosic biomass and release fermentable sugars

Identifieur interne : 001C26 ( Main/Exploration ); précédent : 001C25; suivant : 001C27

Chloroplast‐derived enzyme cocktails hydrolyse lignocellulosic biomass and release fermentable sugars

Auteurs : Dheeraj Verma [États-Unis] ; Anderson Kanagaraj [États-Unis] ; Shuangxia Jin [États-Unis] ; Nameirakpam D. Singh [États-Unis] ; Pappachan E. Kolattukudy [États-Unis] ; Henry Daniell

Source :

Plant Biotechnology Journal [ 1467-7644 ] ; 2010-04.

RBID : ISTEX:A1EC07A0BCBB41E92AD89DD0C75BE0D066E19CAE

English descriptors

KwdEn :
- biofuel, cell wall‐degrading enzymes, cellulosic ethanol, fermentable sugars, lignocellulosic biomass, renewable energy.

Abstract

It is widely recognized that biofuel production from lignocellulosic materials is limited by inadequate technology to efficiently and economically release fermentable sugars from the complex multi‐polymeric raw materials. Therefore, endoglucanases, exoglucanase, pectate lyases, cutinase, swollenin, xylanase, acetyl xylan esterase, beta glucosidase and lipase genes from bacteria or fungi were expressed in Escherichia coli or tobacco chloroplasts. A PCR‐based method was used to clone genes without introns from Trichoderma reesei genomic DNA. Homoplasmic transplastomic lines showed normal phenotype and were fertile. Based on observed expression levels, up to 49, 64 and 10, 751 million units of pectate lyases or endoglucanase can be produced annually, per acre of tobacco. Plant production cost of endoglucanase is 3100‐fold, and pectate lyase is 1057 or 1480‐fold lower than the same recombinant enzymes sold commercially, produced via fermentation. Chloroplast‐derived enzymes had higher temperature stability and wider pH optima than enzymes expressed in E. coli. Plant crude‐extracts showed higher enzyme activity than E. coli with increasing protein concentration, demonstrating their direct utility without purification. Addition of E. coli extracts to the chloroplast‐derived enzymes significantly decreased their activity. Chloroplast‐derived crude‐extract enzyme cocktails yielded more (up to 3625%) glucose from filter paper, pine wood or citrus peel than commercial cocktails. Furthermore, pectate lyase transplastomic plants showed enhanced resistance to Erwina soft rot. This is the first report of using plant‐derived enzyme cocktails for production of fermentable sugars from lignocellulosic biomass. Limitations of higher cost and lower production capacity of fermentation systems are addressed by chloroplast‐derived enzyme cocktails.

Url:

DOI: 10.1111/j.1467-7652.2009.00486.x

Affiliations:

Le document en format XML

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<front><div type="abstract" xml:lang="en">It is widely recognized that biofuel production from lignocellulosic materials is limited by inadequate technology to efficiently and economically release fermentable sugars from the complex multi‐polymeric raw materials. Therefore, endoglucanases, exoglucanase, pectate lyases, cutinase, swollenin, xylanase, acetyl xylan esterase, beta glucosidase and lipase genes from bacteria or fungi were expressed in Escherichia coli or tobacco chloroplasts. A PCR‐based method was used to clone genes without introns from Trichoderma reesei genomic DNA. Homoplasmic transplastomic lines showed normal phenotype and were fertile. Based on observed expression levels, up to 49, 64 and 10, 751 million units of pectate lyases or endoglucanase can be produced annually, per acre of tobacco. Plant production cost of endoglucanase is 3100‐fold, and pectate lyase is 1057 or 1480‐fold lower than the same recombinant enzymes sold commercially, produced via fermentation. Chloroplast‐derived enzymes had higher temperature stability and wider pH optima than enzymes expressed in E. coli. Plant crude‐extracts showed higher enzyme activity than E. coli with increasing protein concentration, demonstrating their direct utility without purification. Addition of E. coli extracts to the chloroplast‐derived enzymes significantly decreased their activity. Chloroplast‐derived crude‐extract enzyme cocktails yielded more (up to 3625%) glucose from filter paper, pine wood or citrus peel than commercial cocktails. Furthermore, pectate lyase transplastomic plants showed enhanced resistance to Erwina soft rot. This is the first report of using plant‐derived enzyme cocktails for production of fermentable sugars from lignocellulosic biomass. Limitations of higher cost and lower production capacity of fermentation systems are addressed by chloroplast‐derived enzyme cocktails.</div>
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Chloroplast‐derived enzyme cocktails hydrolyse lignocellulosic biomass and release fermentable sugars

Chloroplast‐derived enzyme cocktails hydrolyse lignocellulosic biomass and release fermentable sugars

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Abstract

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