title: Genome shuffling improves production of the low-temperature alkalophilic lipase by Acinetobacter johnsonii.
abstract: The production of a low-temperature alkalophilic lipase from Acinetobacter johnsonii was improved using genome shuffling. The starting populations, obtained by UV irradiation and diethyl sulfate mutagenesis, were subjected to recursive protoplast fusion. The optimal conditions for protoplast formation and regeneration were 0.15 mg lysozyme/ml for 45 min at 37°C. The protoplasts were inactivated under UV for 20 min or heated at 60°C for 60 min and a fusant probability of ~98% was observed. The positive colonies were created by fusing the inactivated protoplasts. After two rounds of genome shuffling, one strain, F22, with a lipase activity of 7 U/ml was obtained.
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China. haikuanwangcn@yahoo.com.cn
authors.forename: HaiKuan
authors.initials: H
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: XiaoJie
authors.initials: X
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Qi
authors.forename: YuJie
authors.initials: Y
authors.lastname: Dai
mesh_terms: D000150:Acinetobacter / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects; D003080:Cold Temperature; D032621:DNA Shuffling; D004269:DNA, Bacterial / Q000235:genetics*; D016680:Genome, Bacterial; D008049:Lipase / Q000096:biosynthesis* / Q000235:genetics; D009153:Mutagens / Q000378:metabolism; D011523:Protoplasts; D013463:Sulfuric Acid Esters / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D004269:DNA, Bacterial; D009153:Mutagens; D013463:Sulfuric Acid Esters; D008049:Lipase; C011612:diethyl sulfate