A novel Bacillus ligniniphilus catechol 2,3-dioxygenase shows unique substrate preference and metal requirement

被引：21

作者：

Adewale, Peter ^{[1
]}

Lang, Alice ^{[1
]}

Huang, Fang ^{[1
]}

Zhu, Daochen ^{[2
]}

Sun, Jianzhong ^{[2
]}

Ngadi, Michael ^{[3
]}

Yang, Trent Chunzhong ^{[1
]}

机构：

[1] Natl Res Council Canada, Aquat & Crop Resource Dev Res Ctr, Bioproc & Biocatalysis Team, 100 Sussex Dr, Ottawa, ON K1A 0R6, Canada

[2] Jiangsu Univ, Sch Environm & Safety Engn, Zhenjiang, Jiangsu, Peoples R China

[3] McGill Univ, Bioresource Engn Dept, 21111 Lakeshore Rd, Ste Anne De Bellevue, PQ H9X 3V9, Canada

来源：

SCIENTIFIC REPORTS | 2021年 / 11卷 / 01期

关键词：

PSEUDOMONAS-PUTIDA; EXTRADIOL DIOXYGENASE; 2,3-DIHYDROXYBIPHENYL 1,2-DIOXYGENASE; LIGNIN DEGRADATION; STRAIN; PURIFICATION; ENZYMES; SPECIFICITY; BACTERIA; IRON;

D O I：

10.1038/s41598-021-03144-8

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Identification of novel enzymes from lignin degrading microorganisms will help to develop biotechnologies for biomass valorization and aromatic hydrocarbons degradation. Bacillus ligniniphilus L1 grows with alkaline lignin as the single carbon source and is a great candidate for ligninolytic enzyme identification. The first dioxygenase from strain L1 was heterologously expressed, purified, and characterized with an optimal temperature and pH of 32.5 degrees C and 7.4, respectively. It showed the highest activity with 3-ethylcatechol and significant activities with other substrates in the decreasing order of 3-ethylcatechol > 3-methylcatechol > 3-isopropyl catechol > 2, 3-dihydroxybiphenyl > 4-methylcatechol > catechol. It did not show activities against other tested substrates with similar structures. Most reported catechol 2,3-dioxygenases (C23Os) are Fe2+-dependent whereas Bacillus ligniniphilus catechol 2,3-dioxygenase (BLC23O) is more Mn2+- dependent. At 1 mM, Mn2+ led to 230-fold activity increase and Fe2+ led to 22-fold increase. Sequence comparison and phylogenetic analyses suggested that BL23O is different from other Mn-dependent enzymes and uniquely grouped with an uncharacterized vicinal oxygen chelate (VOC) family protein from Paenibacillus apiaries. Gel filtration analysis showed that BLC23O is a monomer under native condition. This is the first report of a C23O from Bacillus ligniniphilus L1 with unique substrate preference, metal-dependency, and monomeric structure.

引用

页数：16

共 88 条

[1]

ASTURIAS JA, 1994, J BIOL CHEM, V269, P7807

[2] ISOLATION OF A PSEUDOMONAS-STUTZERI STRAIN THAT DEGRADES ORTHO-XYLENE [J].