期刊
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
卷 8, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fbioe.2020.00850
关键词
pectate lyase; high-level expression; biochemical properties; ramie degumming; Pichia pastoris
资金
- National Natural Science Foundation of China [31470519]
- National Science Foundation for Young Scientists of China [31400062]
Pectate lyases play an essential role in textiles, animal feed, and oil extraction industries.Pichia pastoris can be an ideal platform for pectate lyases production, and BspPel (a thermo-alkaline pectate lyase fromBacillussp. RN1) was overexpressed by combined strategies, reaching 1859 U/mL in a 50 L fermentator. It displayed the highest activity at 80 degrees C, and maintained more than 60% of the activity between 30 and 70 degrees C for 1 h. It showed an optimal pH of 10.0, and exhibited remarkable stability over a wider pH range (3.0-11.0), retaining more than 80.0% of enzyme activity for 4 h. TheK(m)andk(cat)of BspPel on PGA (polygalacturonic acid) was 2.19 g L(-1)and 116.1 s(-1), respectively. The activity was significantly enhanced by Ca2+, Mn2+, and Cu2+, and a slight increase was observed with the addition of Ba(2+)and Mg2+. Scanning electron microscope was used to show the degumming efficiency of BspPel on ramie fibers. The loss weight was 9.2% when treated with crude enzyme supernatant and 20.8% when treated with the enzyme-chemical method, which was higher than the 14.2% weight loss in the positive control treated with 0.5% (w/v) NaOH alone. In conclusion, BspPel could be a good candidate for the ramie degumming industry.
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