Improving the thermoactivity and thermostability of pectate lyase from Bacillus pumilus for ramie degumming

Verfasser / Beitragende:
[Chaoning Liang, Xiwu Gui, Cheng Zhou, Yanfen Xue, Yanhe Ma, Shuang-Yan Tang]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/6(2015-03-01), 2673-2682
Format:
Artikel (online)
ID: 605499055
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024 7 0 |a 10.1007/s00253-014-6091-y  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-014-6091-y 
245 0 0 |a Improving the thermoactivity and thermostability of pectate lyase from Bacillus pumilus for ramie degumming  |h [Elektronische Daten]  |c [Chaoning Liang, Xiwu Gui, Cheng Zhou, Yanfen Xue, Yanhe Ma, Shuang-Yan Tang] 
520 3 |a 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. 
540 |a Springer-Verlag Berlin Heidelberg, 2014 
690 7 |a Pectate lyase  |2 nationallicence 
690 7 |a Directed evolution  |2 nationallicence 
690 7 |a Thermostability  |2 nationallicence 
690 7 |a Ramie degumming  |2 nationallicence 
700 1 |a Liang  |D Chaoning  |u 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  |4 aut 
700 1 |a Gui  |D Xiwu  |u College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, 030801, Taigu, Shanxi, People's Republic of China  |4 aut 
700 1 |a Zhou  |D Cheng  |u 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  |4 aut 
700 1 |a Xue  |D Yanfen  |u 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  |4 aut 
700 1 |a Ma  |D Yanhe  |u 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  |4 aut 
700 1 |a Tang  |D Shuang-Yan  |u 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  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/6(2015-03-01), 2673-2682  |x 0175-7598  |q 99:6<2673  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-014-6091-y  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-014-6091-y  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Liang  |D Chaoning  |u 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  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Gui  |D Xiwu  |u College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, 030801, Taigu, Shanxi, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Zhou  |D Cheng  |u 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  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Xue  |D Yanfen  |u 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  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Ma  |D Yanhe  |u 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  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Tang  |D Shuang-Yan  |u 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  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/6(2015-03-01), 2673-2682  |x 0175-7598  |q 99:6<2673  |1 2015  |2 99  |o 253