Archives of Biological Sciences 2013 Volume 65, Issue 1, Pages: 305-314
https://doi.org/10.2298/ABS1301305Q
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Improved cellulolytic efficacy in Penicilium decumbens via heterologous expression of Hypocrea jecorina endoglucanase II
Qin Yuqi (National Glycoengineering Research Center, Shandong University, Jinan, China)
Zheng Kai (State Key Laboratory of Microbial Technology, Shandong University, Jinan, China)
Liu Guodong (State Key Laboratory of Microbial Technology, Shandong University, Jinan, China)
Chen Mei (State Key Laboratory of Microbial Technology, Shandong University, Jinan, China)
Qu Yinbo (National Glycoengineering Research Center, Shandong University, Jinan, China + State Key Laboratory of Microbial Technology, Shandong University, Jinan, China)
Hypocrea jecorina endoglucanase II (Hjegl2) was heterologously expressed in
Penicillium decumbens (yielding strain Pd::Hjegl2). After induction in
cellulose containing media, strain Pd::Hjeg2 displayed increased
carboxymethylcellulase activity (CMCase, 5.77 IU/ml, representing a 21%
increase) and cellulose degradation determined with a filter paper assay
(FPA, 0.40 IU/ml, 67% increase), as compared to the parent strain. In media
supplemented with glucose (2%), Pd::Hjegl2, displayed 51.2-fold and 3-fold
higher CMCase and FPA activities, respectively, as compared to the parent
strain. No changes in the expression levels of the four main native cellulase
genes of P. decumbens (Pdegl1, Pdegl2, Pdcbh1, and Pdcbh2) were noted between
the transformant and wild-type strains. These data support the idea that
Hjegl2 cleaves both internal and terminal glycosidic residues, in a
relatively random and processive manner. In situ polyacrylamide gelactivity
staining of extracts derived from wild-type and Pd::Hjegl2 revealed two
additional active fractions in the latter strain; one with a molecular mass
~50-65 KDa and another ~80-116 kDa.
Keywords: Penicillium decumbens, endoglucanase II, cellulose degradation, heterologous expression