An L-arabinose isomerase from Acidothermus cellulolytics ATCC 43068: cloning, expression, purification, and characterization

Applied Microbiology and Biotechnology
Lifang ChengBo Jiang

Abstract

The araA gene encoding an L-arabinose isomerase (L-AI) from the acido-thermophilic bacterium Acidothermus cellulolytics ATCC 43068 was cloned and overexpressed in Escherichia coli. The open reading frame of the L-AI consisted of 1,503 nucleotides encoding 501 amino acid residues. The recombinant L-AI was purified to homogeneity by heat treatment, ion-exchange chromatography, and gel filtration. The molecular mass of the enzyme was estimated to be approximately 55 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified enzyme was optimally active at 75 degrees C and pH 7.5. It required divalent metal ions, either Mn(2+) or Co(2+), for both enzymatic activity and thermostability improvement at higher temperatures. The enzyme showed relatively high activity and stability at acidic pH. It exhibited over 90% of its maximal activity at pH 6.0 and retained 80% of activity after 12 h incubation at pH 6.0. Catalytic property study showed that the enzyme had an interesting catalytic efficiency. Its apparent K (m), V (max), and catalytic efficiency (k (cat)/K (m)) for D-galactose was 28.9 mM, 4.9 U/mg, and 9.3 mM(-1) min(-1), respectively. The enzyme carried out the isomerization of D-galactose to D-tagatose with a ...Continue Reading

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Citations

May 29, 2014·Applied Microbiology and Biotechnology·Wanmeng MuBo Jiang
Oct 5, 2014·Applied Microbiology and Biotechnology·Zheng XuHong Xu
Jul 19, 2012·World Journal of Microbiology & Biotechnology·Xingding Zhou, Jin Chuan Wu
Feb 22, 2012·Journal of Industrial Microbiology & Biotechnology·Koen BeerensWim Soetaert
Jan 9, 2016·Critical Reviews in Food Science and Nutrition·J JayamuthunagaiM Chakravarthy
Jul 8, 2015·Applied Microbiology and Biotechnology·Wanmeng MuBo Jiang
May 23, 2017·Journal of Agricultural and Food Chemistry·Zhaojuan ZhengJia Ouyang
Oct 20, 2018·Journal of Food Science·Sohini RoyAshis Kumar Samanta
Feb 9, 2013·Bioscience, Biotechnology, and Biochemistry·Myung-Ji Seo
Aug 20, 2016·Applied Biochemistry and Biotechnology·Shubo LiZongwen Pang
Feb 6, 2020·Bioprocess and Biosystems Engineering·Chengzhu KeZongwen Pang
May 16, 2020·Frontiers in Bioengineering and Biotechnology·Guoyan ZhangXianghui Qi
Nov 16, 2018·Applied Biochemistry and Biotechnology·Marylane de SousaLuciana R B Gonçalves
Nov 23, 2011·Journal of Agricultural and Food Chemistry·Young-Ho HongSung Haeng Lee

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