caa a22 4500 605499055 CHVBK 20210128100549.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6091-y doi (NATIONALLICENCE)springer-10.1007/s00253-014-6091-y Improving the thermoactivity and thermostability of pectate lyase from Bacillus pumilus for ramie degumming [Elektronische Daten] [Chaoning Liang, Xiwu Gui, Cheng Zhou, Yanfen Xue, Yanhe Ma, Shuang-Yan Tang] Thermostable alkaline pectate lyases can be potentially used for enzymatically degumming ramie in an environmentally sustainable manner and as an alternative to the currently used chemical-based ramie degumming processes. To assess its potential applications, pectate lyase from Bacillus pumilus (ATCC 7061) was cloned and expressed in Escherichia coli. Evolutionary strategies were applied to generate efficient ramie degumming enzymes. Obtained from site-saturation mutagenesis and random mutagenesis, the best performing mutant enzyme M3 exhibited a 3.4-fold higher specific activity on substrate polygalacturonic acid, compared with the wild-type enzyme. Furthermore, the half-life of inactivation at 50°C for M3 mutant extended to over 13h. In contrast, the wild-type enzyme was completely inactivated in less than 10min under the same conditions. An upward shift in the optimal reaction temperature of M3 mutant, to 75°C, was observed, which was 10°C higher than that of the wild-type enzyme. Kinetic parameter data revealed that the catalysis efficiency of M3 mutant was higher than that of the wild-type enzyme. Ramie degumming with M3 mutant was also demonstrated to be more efficient than that with the wild-type enzyme. Collectively, our results suggest that the M3 mutant, with remarkable improvements in thermoactivity and thermostability, has potential applications for ramie degumming in the textile industry. Springer-Verlag Berlin Heidelberg, 2014 Pectate lyase nationallicence Directed evolution nationallicence Thermostability nationallicence Ramie degumming nationallicence Liang Chaoning CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut Gui Xiwu College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, 030801, Taigu, Shanxi, People's Republic of China aut Zhou Cheng State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut Xue Yanfen State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut Ma Yanhe State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut Tang Shuang-Yan CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2673-2682 0175-7598 99:6<2673 2015 99 253 https://doi.org/10.1007/s00253-014-6091-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6091-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liang Chaoning CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Gui Xiwu College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, 030801, Taigu, Shanxi, People's Republic of China aut NATIONALLICENCE 700 1- Zhou Cheng State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Xue Yanfen State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Ma Yanhe State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Tang Shuang-Yan CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, 100101, Chaoyang District, Beijing, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2673-2682 0175-7598 99:6<2673 2015 99 253