Mixture optimization of six core glycosyl hydrolases for maximizing saccharification of ammonia fiber expansion (AFEX) pretreated corn stover

Dahai Gao, Shishir P.S. Chundawat, Chandraraj Krishnan, Venkatesh Balan, Bruce E. Dale

Research output: Contribution to journalArticlepeer-review

164 Scopus citations

Abstract

In this work, six core glycosyl hydrolases (GH) were isolated and purified from various sources to help rationally optimize an enzyme cocktail to digest ammonia fiber expansion (AFEX) treated corn stover. The four core cellulases were endoglucanase I (EG I, GH family 7B), cellobiohydrolase I (CBH I, GH family 7A), cellobiohydrolase II (CBH II, GH family 6A) and β-glucosidase (βG, GH family 3). The two core hemicellulases were an endo-xylanase (EX, GH family 11) and a β-xylosidase (βX, GH family 3). Enzyme family and purity were confirmed by proteomics. Synergistic interactions among the six core enzymes for varying relative and total protein loading (8.25, 16.5 and 33 mg/g glucan) during hydrolysis of AFEX-treated corn stover was studied using a high-throughput microplate based protocol. The optimal composition (based on% protein mass loading) of the cocktail mixture was CBH I (28.4%): CBH II (18.0%): EG I (31.0%): EX (14.1%): βG (4.7%): βX (3.8%). These results demonstrate a rational strategy for the development of a minimal, synergistic enzymes cocktail that could reduce enzyme usage and maximize the fermentable sugar yields from pretreated lignocellulosics.

Original languageEnglish (US)
Pages (from-to)2770-2781
Number of pages12
JournalBioresource Technology
Volume101
Issue number8
DOIs
StatePublished - Apr 2010
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Bioengineering
  • Environmental Engineering
  • Renewable Energy, Sustainability and the Environment
  • Waste Management and Disposal

Keywords

  • AFEX pretreatment
  • Cellulase
  • Cellulosic ethanol
  • Enzymatic hydrolysis
  • Hemicellulase

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