Heterologous expression of cellobiohydrolase II (Cel6A) in maize endosperm

• Published in: Transgenic research Vol. 22; no. 3; pp. 477 - 488
• Main Authors: Devaiah, Shivakumar Pattada, Requesens, Deborah Vicuna, Chang, Yeun-Kyung, Hood, Kendall R, Flory, Ashley, Howard, John A, Hood, Elizabeth E
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer-Verlag 01.06.2013; Springer Netherlands; Springer; Springer Nature B.V
• Subjects: Animal Genetics and Genomics; biofuels; Biological and medical sciences; biomass; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology; biotransformation; cellulose; Cellulose - metabolism; cellulose 1,4-beta-cellobiosidase; Cellulose 1,4-beta-Cellobiosidase - genetics; Cellulose 1,4-beta-Cellobiosidase - isolation & purification; Cellulose 1,4-beta-Cellobiosidase - metabolism; corn; endo-1,4-beta-glucanase; endosperm; Endosperm - enzymology; Endosperm - genetics; enzyme activity; Fundamental and applied biological sciences. Psychology; Genetic Engineering; Genetic technics; Hypocrea - enzymology; Hypocrea - genetics; Hypocrea jecorina; Life Sciences; Methods. Procedures. Technologies; microbial growth; Molecular Medicine; Original Paper; Plant Genetics and Genomics; Plants, Genetically Modified - genetics; Plants, Genetically Modified - growth & development; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Saccharomyces cerevisiae - growth & development; seeds; Transgenic animals and transgenic plants; Transgenics; Zea mays; Zea mays - genetics; Zea mays - growth & development; Endosperm; Cel6A; Cellobiohydrolase II; Biomass; Hypocrea jecorina; Monocotyledones; Zea mays; Enzyme; Cellulose 1,4-β-cellobiosidase; Gene expression; Glycosylases; Gramineae; Angiospermae; Hydrolases; Spermatophyta
• ISSN: 0962-8819; 1573-9368
• DOI: 10.1007/s11248-012-9659-2

The technology of converting lignocellulose to biofuels has advanced swiftly over the past few years, and enzymes are a significant constituent of this technology. In this regard, cost effective production of cellulases has been the focus of research for many years. One approach to reach cost targets of these enzymes involves the use of plants as bio-factories. The application of this technology to plant biomass conversion for biofuels and biobased products has the potential for significantly lowering the cost of these products due to lower enzyme production costs. Cel6A, one of the two cellobiohydrolases (CBH II) produced by Hypocrea jecorina, is an exoglucanase that cleaves primarily cellobiose units from the non-reducing end of cellulose microfibrils. In this work we describe the expression of Cel6A in maize endosperm as part of the process to lower the cost of this dominant enzyme for the bioconversion process. The enzyme is active on microcrystalline cellulose as exponential microbial growth was observed in the mixture of cellulose, cellulases, yeast and Cel6A, Cel7A (endoglucanase), and Cel5A (cellobiohydrolase I) expressed in maize seeds. We quantify the amount accumulated and the activity of the enzyme. Cel6A expressed in maize endosperm was purified to homogeneity and verified using peptide mass finger printing.