Expression and Extracellular Secretion of Endo-glucanase and Xylanase by Zymomonas mobilis

Autor: Verawat Champreda, Apinya Sowatad, Phattara-orn Havanapan, Pattanop Kanokratana, Tatsaporn Todhanakasem
Rok vydání: 2018
Předmět:
Zdroj: Applied Biochemistry and Biotechnology. 187:239-252
ISSN: 1559-0291
0273-2289
DOI: 10.1007/s12010-018-2821-4
Popis: Recombinant Zymomonas mobilis (pGEX-4T-3 BI 120-2) was constructed to encode endo-glucanase (CelA) and endo-xylanase (Xyn11) from Z. mobilis ZM4 (ATCC 31821) and an uncultured bacterium. The recombinant was genetically engineered with the N-terminus of a predicted SecB-dependent (type II) secretion signal from phoC of Z. mobilis to translocate the enzymes extracellularly. Both the enzymes were characterized regarding their functional optimum pH and temperature, with the highest multi-enzyme activities at pH 6.0 and a temperature of 30 °C, which approximates the optimum conditions for ethanol production by Z. mobilis. The crude intracellular and extracellular fractions of the recombinant were characterized in terms of substrate specificity using carboxymethyl cellulose (CMC), beechwood xylan, filter paper, Avicel, and pretreated rice straw. The crude extracellular and intracellular enzymes with cellulolytic and xylanolytic activities were more robustly produced and secreted from the recombinant strain compared to the wild-type and ampicillin-sensitive strains, using CMC and beechwood xylan as the substrates. Ethanol production by the recombinant strain was greater than the production by the wild-type strain when pretreated rice straw was used as a sole carbon source.
Databáze: OpenAIRE
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