caa a22 4500 606213244 CHVBK 20210128101046.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s10295-014-1571-8 doi (NATIONALLICENCE)springer-10.1007/s10295-014-1571-8 Cloning and expression of a novel prolyl endopeptidase from Aspergillus oryzae and its application in beer stabilization [Elektronische Daten] [Chao Kang, Xiao-Wei Yu, Yan Xu] A novel prolyl endopeptidase gene from Aspergillus oryzae was cloned and expressed in Pichia pastoris. Amino acid sequence analysis of the prolyl endopeptidase from Aspergillus oryzae (AO-PEP) showed that this enzyme belongs to a class serine peptide S28 family. Expression, purification and characterization of AO-PEP were analyzed. The optimum pH and temperature were pH 5.0 and 40°C, respectively. The enzyme was activated and stabilized by metal ion Ca2+ and inhibited by Zn2+, Mn2+, Al3+, and Cu2+. The K m and k cat values of the purified enzyme for different substrates were evaluated. The results implied that the recombinant AO-PEP possessed higher affinity for the larger substrate. A fed-batch strategy was developed for the high-cell-density fermentation and the enzyme activity reached 1,130U/l after cultivation in 7l fermentor. After addition of AO-PEP during the fermentation phase of beer brewing, demonstrated the potential application of AO-PEP in the non-biological stability of beer, which favor further industrial development of this new enzyme in beer stabilization, due to its reducing operational costs, as well as no beer losses unlike regeneration process and beer lost with regenerated polyvinylpolypyrrolidone system. Society for Industrial Microbiology and Biotechnology, 2014 Prolyl endopeptidase nationallicence Aspergillus oryzae nationallicence Cloning and expression nationallicence Beer stabilization nationallicence Kang Chao The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, 214122, Wuxi, Jiangsu, China aut Yu Xiao-Wei The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, 214122, Wuxi, Jiangsu, China aut Xu Yan The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, 214122, Wuxi, Jiangsu, China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/2(2015-02-01), 263-272 1367-5435 42:2<263 2015 42 10295 https://doi.org/10.1007/s10295-014-1571-8 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/s10295-014-1571-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kang Chao The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, 214122, Wuxi, Jiangsu, China aut NATIONALLICENCE 700 1- Yu Xiao-Wei The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, 214122, Wuxi, Jiangsu, China aut NATIONALLICENCE 700 1- Xu Yan The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, 214122, Wuxi, Jiangsu, China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/2(2015-02-01), 263-272 1367-5435 42:2<263 2015 42 10295