Protein engineering of the aldoxime dehydratase from Bacillus sp. OxB-1 based on a rational sequence alignment approach

被引：8

作者：

Oike, Keiko ^{[1
]}

Spross, Jens ^{[1
]}

Matsui, Daisuke ^{[2
,3
]}

Asano, Yasuhisa ^{[2
,3
]}

Groeger, Harald ^{[1
]}

机构：

[1] Bielefeld Univ, Fac Chem, Chair Ind Organ Chem & Biotechnol, Univ Str 25, D-33615 Bielefeld, Germany

[2] Toyama Prefectural Univ, Biotechnol Res Ctr, 5180 Kurokawa, Imizu, Toyama 9390398, Japan

[3] Toyama Prefectural Univ, Dept Biotechnol, 5180 Kurokawa, Imizu, Toyama 9390398, Japan

来源：

SCIENTIFIC REPORTS | 2021年 / 11卷 / 01期

关键词：

HEME-CONTAINING LYASE; TRIPLE BOND SYNTHESIS; CYANIDE-FREE; PHENYLACETALDOXIME DEHYDRATASE; NITRILE HYDRATASE; ESCHERICHIA-COLI; STRAIN OXB-1; PURIFICATION; EXPRESSION; IDENTIFICATION;

D O I：

10.1038/s41598-021-92749-0

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：

Recently, the program INTMSAlign_HiSol for identifying aggregation hotspots in proteins only requiring secondary structure data was introduced. We explored the utility of this program further and applied it for engineering of the aldoxime dehydratase from Bacillus sp. OxB-1. Towards this end, the effect of inverting the hydropathy at selected positions of the amino acid sequence on the enzymatic activity was studied leading to 60% of our constructed variants, which showed improved activity. In part, this activity increase can be rationalised by an improved heme incorporation of the variants. For example, a single mutation gave a 1.8 fold increased enzymatic activity and 30% improved absolute heme incorporation.

引用

页数：13

共 30 条

[1] Protein engineering of the aldoxime dehydratase from Bacillus sp. OxB-1 based on a rational sequence alignment approach
Keiko Oike
Jens Sproß
Daisuke Matsui
Yasuhisa Asano
Harald Gröger
Scientific Reports, 11
[2] Complete Genome Sequence of an Aldoxime Degrader, Bacillus sp. OxB-1
Yamaguchi, Takuya
Asano, Yasuhisa
GENOME ANNOUNCEMENTS, 2015, 3 (01)
[3] Z-phenylacetaldoxime degradation by a novel aldoxime dehydratase from Bacillus sp. strain OxB-1
Asano, Y
Kato, Y
FEMS MICROBIOLOGY LETTERS, 1998, 158 (02) : 185 - 190
[4] Rapid purification of aldoxime dehydratase from Bacillus sp OxB-1 and spectroscopic analysis of the enzyme
不详
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 2005, 36 (1-6) : 58 - 58
[5] Crystal structural analysis of aldoxime dehydratase from Bacillus sp. OxB-1: Importance of surface residues in optimization for crystallization
Matsui, Daisuke
Muraki, Norifumi
Chen, Ke
Mori, Tomoya
Ingram, Aaron A.
Oike, Keiko
Groger, Harald
Aono, Shigetoshi
Asano, Yasuhisa
JOURNAL OF INORGANIC BIOCHEMISTRY, 2022, 230
[6] High yield synthesis of nitriles by a new enzyme, phenylacetaldoxime dehydratase, from Bacillus sp strain OxB-1
Xie, SX
Kato, Y
Asano, Y
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2001, 65 (12) : 2666 - 2672
[7] Novel heme-containing lyase, phenylacetaldoxime dehydratase from Bacillus sp, strain OxB-1:: Purification, characterization, and molecular cloning of the gene
Kato, Y
Nakamura, K
Sakiyama, H
Mayhew, SG
Asano, Y
BIOCHEMISTRY, 2000, 39 (04) : 800 - 809
[8] Purification and partial characterization of N-hydroxy-L-phenylalanine decarboxylase/oxidase from Bacillus sp strain OxB-1, an enzyme involved in aldoxime biosynthesis in the "aldoxime-nitrile pathway"
Kato, Yasuo
Tsuda, Takashi
Asano, Yasuhisa
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 2007, 1774 (07): : 856 - 865
[9] High-level expression of a novel FMN-dependent heme-containing lyase, phenylacetaldoxime dehydratase of Bacillus sp strain OxB-1, in heterologous hosts
Kato, Y
Asano, Y
PROTEIN EXPRESSION AND PURIFICATION, 2003, 28 (01) : 131 - 139
[10] In Silico Rational Design and Protein Engineering of Disulfide Bridges of an α-Amylase from Geobacillus sp. to Improve Thermostability
Zhang, Heng
Zhai, Wenxin
Lin, Lin
Wang, Ping
Xu, Xiangyang
Wei, Wei
Wei, Dongzhi
STARCH-STARKE, 2021, 73 (9-10):

← 1 2 3 →