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通过对米曲霉中参与碳分解代谢物阻遏的creA和creB基因进行双缺失来提高生物质降解酶的产量。

Increased production of biomass-degrading enzymes by double deletion of creA and creB genes involved in carbon catabolite repression in Aspergillus oryzae.

作者信息

Ichinose Sakurako, Tanaka Mizuki, Shintani Takahiro, Gomi Katsuya

机构信息

Laboratory of Bioindustrial Genomics, Graduate School of Agricultural Science, Tohoku University, 468-1 Aoba, Aramaki, Aoba-ku, Sendai 980-0845, Japan.

Laboratory of Bioindustrial Genomics, Graduate School of Agricultural Science, Tohoku University, 468-1 Aoba, Aramaki, Aoba-ku, Sendai 980-0845, Japan.

出版信息

J Biosci Bioeng. 2018 Feb;125(2):141-147. doi: 10.1016/j.jbiosc.2017.08.019. Epub 2017 Sep 30.

DOI:10.1016/j.jbiosc.2017.08.019
PMID:28970110
Abstract

In a previous study, we reported that a double gene deletion mutant for CreA and CreB, which constitute the regulatory machinery involved in carbon catabolite repression, exhibited improved production of α-amylase compared with the wild-type strain and single creA or creB deletion mutants in Aspergillus oryzae. Because A. oryzae can also produce biomass-degrading enzymes, such as xylolytic and cellulolytic enzymes, we examined the production levels of those enzymes in deletion mutants in this study. Xylanase and β-glucosidase activities in the wild-type were hardly detected in submerged culture containing xylose as the carbon source, whereas those enzyme activities were significantly increased in the single creA deletion (ΔcreA) and double creA and creB deletion (ΔcreAΔcreB) mutants. In particular, the ΔcreAΔcreB mutant exhibited >100-fold higher xylanase and β-glucosidase activities than the wild-type. Moreover, in solid-state culture, the β-glucosidase activity of the double deletion mutant was >7-fold higher than in the wild-type. These results suggested that deletion of both creA and creB genes could also efficiently improve the production levels of biomass-degrading enzymes in A. oryzae.

摘要

在之前的一项研究中,我们报道了构成碳分解代谢物阻遏调控机制的CreA和CreB双基因缺失突变体,与米曲霉中的野生型菌株以及单一的creA或creB缺失突变体相比,其α-淀粉酶产量有所提高。由于米曲霉还能产生生物质降解酶,如木聚糖分解酶和纤维素分解酶,因此在本研究中我们检测了这些酶在缺失突变体中的产量水平。在以木糖作为碳源的深层培养中,野生型中的木聚糖酶和β-葡萄糖苷酶活性几乎检测不到,而在单一的creA缺失(ΔcreA)以及creA和creB双缺失(ΔcreAΔcreB)突变体中,这些酶的活性显著增加。特别是,ΔcreAΔcreB突变体的木聚糖酶和β-葡萄糖苷酶活性比野生型高100倍以上。此外,在固态培养中,双缺失突变体的β-葡萄糖苷酶活性比野生型高7倍以上。这些结果表明,creA和creB基因的缺失也能有效提高米曲霉中生物质降解酶的产量水平。

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