title: Heterologous production of an acidic thermostable lipase with broad-range pH activity from thermophilic fungus Neosartorya fischeri P1.
abstract: Thermophilic Neosartorya fischeri P1 is an excellent lipase producer and harbors seven lipase genes. All genes were found to be functional after heterologous expression in Escherichia coli. One of them, LIP09, showed high-level expression in Pichia pastoris with the yield of 2.0 g/L in a 3.7-L fermentor. Deduced amino acid sequence of LIP09 consists of a putative signal peptide (residues 1-19) and a mature polypeptide (residues 20-562). Compared with other fungal counterparts, purified recombinant LIP09 has some superior properties. It exhibited maximum activity at 60°C and pH 5.0, had broad pH adaptability (>60% activity at pH 3.5-8.0) and stability (retaining >90% activity after incubation at pH 3.0-7.0 for 1 h at 40°C), and was highly thermostable (retaining >96% activity after incubation at 50°C for 30 min). The r-LIP09 had a preference for the medium-chain length p-nitrophenyl esters (C12) rather than short and long-chain length substrates. The high-level expression and excellent properties make LIP09 a potential enzyme candidate in food and feed industries.
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China; Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Bioengineering, Tianjin University of Science & Technology, Tianjin 300457, PR China.
authors: Qiaoqiao
authors: Q
authors: Sun
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.
authors: Hui
authors: H
authors: Wang
authors: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Bioengineering, Tianjin University of Science & Technology, Tianjin 300457, PR China.
authors: Huitu
authors: H
authors: Zhang
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.
authors: Huiying
authors: H
authors: Luo
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.
authors: Pengjun
authors: P
authors: Shi
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.
authors: Yingguo
authors: Y
authors: Bai
authors: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Bioengineering, Tianjin University of Science & Technology, Tianjin 300457, PR China.
authors: Fuping
authors: F
authors: Lu
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.
authors: Bin
authors: B
authors: Yao
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China. Electronic address: huanghuoqing@caas.cn.
authors: Huoqing
authors: H
authors: Huang
mesh_terms: D000220:Adaptation, Biological / Q000235:genetics; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D004795:Enzyme Stability / Q000235:genetics; D004926:Escherichia coli; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000096:biosynthesis* / Q000235:genetics; D055326:Neosartorya / Q000201:enzymology* / Q000235:genetics*; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D008049:Lipase; C428386:thermostable lipase
keywords: Broad pH adaptability; Lipase; Neosartorya fischeri; Pichia pastoris; Thermophilic fungus