Product inhibition of cellulases studied with 14C-labeled cellulose substrates

Biotechnology for Biofuels

Hele Teugjas¹  Priit Väljamäe¹ 

[1] Institute of Molecular and Cell Biology, University of Tartu, Riia 23b – 202, Tartu 51010, Estonia

关键词: Trichoderma reesei;    Chaetomium thermophilum;    Thermoascus aurantiacus;    Acremonium thermophilum;    Inhibition;    Glucose;    Cellobiose;    Cellulose;    Cellulase;   

Others  :  797972
DOI  :  10.1186/1754-6834-6-104

 received in 2013-05-20, accepted in 2013-07-11,  发布年份 2013

【 摘 要 】

Background

As a green alternative for the production of transportation fuels, the enzymatic hydrolysis of lignocellulose and subsequent fermentation to ethanol are being intensively researched. To be economically feasible, the hydrolysis of lignocellulose must be conducted at a high concentration of solids, which results in high concentrations of hydrolysis end-products, cellobiose and glucose, making the relief of product inhibition of cellulases a major challenge in the process. However, little quantitative information on the product inhibition of individual cellulases acting on cellulose substrates is available because it is experimentally difficult to assess the hydrolysis of the heterogeneous polymeric substrate in the high background of added products.

Results

The cellobiose and glucose inhibition of thermostable cellulases from Acremonium thermophilum, Thermoascus aurantiacus, and Chaetomium thermophilum acting on uniformly ¹⁴C-labeled bacterial cellulose and its derivatives, ¹⁴C-bacterial microcrystalline cellulose and ¹⁴C-amorphous cellulose, was studied. Cellulases from Trichoderma reesei were used for comparison. The enzymes most sensitive to cellobiose inhibition were glycoside hydrolase (GH) family 7 cellobiohydrolases (CBHs), followed by family 6 CBHs and endoglucanases (EGs). The strength of glucose inhibition followed the same order. The product inhibition of all enzymes was relieved at higher temperatures. The inhibition strength measured for GH7 CBHs with low molecular-weight model substrates did not correlate with that measured with ¹⁴C-cellulose substrates.

Conclusions

GH7 CBHs are the primary targets for product inhibition of the synergistic hydrolysis of cellulose. The inhibition must be studied on cellulose substrates instead of on low molecular-weight model substrates when selecting enzymes for lignocellulose hydrolysis. The advantages of using higher temperatures are an increase in the catalytic efficiency of enzymes and the relief of product inhibition.

【 授权许可】

   
2013 Teugjas and Väljamäe; licensee BioMed Central Ltd.

【 图 表 】

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