title: Genome shuffling enhances lipase production of thermophilic Geobacillus sp.
abstract: Thermostable lipases are potential enzymes for biocatalytic application. In this study, the lipase production of Geobacillus sp. CF03 (WT) was improved by genome shuffling. After two rounds of genome shuffling, one fusant strain (FB1) achieved increase lipase activity from the populations generated by ultraviolet irradiation and ethyl methylsulfonate (EMS) mutagenesis. The growth rate and lipase production of FB1 increased highest by 150 and 238 %, respectively, in comparison to the wild type. The fusant enzyme had a significant change in substrate specificity but still prefers the long-chain length substrates. It had an optimum activity at 60 °C, pH at 7.0-8.0, with p-nitrophenyl palmitate (C16) as a substrate and retained about 50 % of their activity after 15 min at 70 °C, pH 8.0. Furthermore, the fusant lipase showed the preference of sesame oil, waste palm oil, and canola oil. Therefore, the genome shuffling strategy has been successful to strain improvement and selecting strain with multiple desirable characteristics.
authors.affiliation: Department of Biochemistry, Faculty of Science, Kasetsart University, 50 Ngam Wong Wan Rd, Chatuchak, Bangkok, 10900, Thailand.
authors.forename: Pornchanok
authors.initials: P
authors.lastname: Chalopagorn
authors.affiliation: Department of Biochemistry and Centre of Excellence for Innovation in Chemistry (PERCH-CIC), Faculty of Science, Burapha University, Bangsaen, Chonburi, 20131, Thailand. jittima@buu.ac.th.|Environmental Science Program, Faculty of Science, Burapha University, Bangsaen, Chonburi, 20131, Thailand. jittima@buu.ac.th.|Center of Excellence on Environmental Health and Toxicology (CHE), Ministry of Education, Bangkok, Thailand. jittima@buu.ac.th.
authors.forename: Jittima
authors.initials: J
authors.lastname: Charoenpanich
authors.affiliation: Department of Biochemistry, Faculty of Science, Kasetsart University, 50 Ngam Wong Wan Rd, Chatuchak, Bangkok, 10900, Thailand. fsciktc@ku.ac.th.|Center for Advanced Studies in Tropical Natural Resources, National Research University-Kasetsart University, Thailand CASTNAR, NRU-KU, Bangkok, Thailand. fsciktc@ku.ac.th.
authors.forename: Kiattawee
authors.initials: K
authors.lastname: Choowongkomon
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D032621:DNA Shuffling; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D016680:Genome, Bacterial; D056506:Geobacillus / Q000201:enzymology / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D000073878:Palm Oil; D010938:Plant Oils / Q000737:chemistry; D000074262:Rapeseed Oil; D012715:Sesame Oil / Q000737:chemistry; D014466:Ultraviolet Rays
chemical_list: D001426:Bacterial Proteins; D005229:Fatty Acids, Monounsaturated; D010938:Plant Oils; D000074262:Rapeseed Oil; D000073878:Palm Oil; D012715:Sesame Oil; D008049:Lipase