A novel low-temperature active alkaline pectate lyase from Klebsiella sp. Y1 with potential in textile industry

2011 ◽  
Vol 46 (10) ◽  
pp. 1921-1926 ◽  
Author(s):  
Peng Yuan ◽  
Kun Meng ◽  
Huiying Luo ◽  
Pengjun Shi ◽  
Huoqing Huang ◽  
...  
Keyword(s):  
Low Temperature ◽  
Textile Industry ◽  
Pectate Lyase ◽  
Alkaline Pectate Lyase

scholarly journals Directed Evolution and Structural Analysis of Alkaline Pectate Lyase from the Alkaliphilic Bacterium Bacillus sp. Strain N16-5 To Improve Its Thermostability for Efficient Ramie Degumming

2015 ◽  
Vol 81 (17) ◽  
pp. 5714-5723 ◽  
Author(s):  
Cheng Zhou ◽  
Jintong Ye ◽  
Yanfen Xue ◽  
Yanhe Ma
Keyword(s):  
Directed Evolution ◽  
Textile Industry ◽  
Specific Activity ◽  
High Specific Activity ◽  
Pectate Lyase ◽  
Fold Increase ◽  
Site Directed Mutagenesis ◽  
Wild Type ◽  
Content Type ◽  
Alkaline Pectate Lyase

ABSTRACTThermostable alkaline pectate lyases have potential applications in the textile industry as an alternative to chemical-based ramie degumming processes. In particular, the alkaline pectate lyase fromBacillussp. strain N16-5 (BspPelA) has potential for enzymatic ramie degumming because of its high specific activity under extremely alkaline conditions without the requirement for additional Ca2+. However, BspPelA displays poor thermostability and is inactive after incubation at 50°C for only 30 min. Here, directed evolution was used to improve the thermostability of BspPelA for efficient and stable degumming. After two rounds of error-prone PCR and screening of >12,000 mutants, 10 mutants with improved thermostability were obtained. Sequence analysis and site-directed mutagenesis revealed that single E124I, T178A, and S271G substitutions were responsible for improving thermostability. Structural and molecular dynamic simulation analysis indicated that the formation of a hydrophobic cluster and new H-bond networks was the key factor contributing to the improvement in thermostability with these three substitutions. The most thermostable combined mutant, EAET, exhibited a 140-fold increase in thet50(time at which the enzyme loses 50% of its initial activity) value at 50°C, accompanied by an 84.3% decrease in activity compared with that of wild-type BspPelA, while the most advantageous combined mutant, EA, exhibited a 24-fold increase in thet50value at 50°C, with a 23.3% increase in activity. Ramie degumming with the EA mutant was more efficient than that with wild-type BspPelA. Collectively, our results suggest that the EA mutant, exhibiting remarkable improvements in thermostability and activity, has the potential for applications in ramie degumming in the textile industry.

scholarly journals Screening of a Novel Polysaccharide Lyase Family 10 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression and Characterization

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 2774 ◽  
Author(s):  
Yan Zhao ◽  
Ye Yuan ◽  
Xinyu Zhang ◽  
Yumei Li ◽  
Qiang Li ◽  
...  
Keyword(s):  
Textile Industry ◽  
Pectate Lyase ◽  
Paenibacillus Polymyxa ◽  
Polygalacturonic Acid ◽  
Optimal Conditions ◽  
Citrus Pectin ◽  
Plant Fiber ◽  
Polysaccharide Lyase ◽  
Glycosidic Bonds ◽  
Fiber Processing

Pectate lyase (EC 4.2.2.2) catalyzes the cleavage of α-1,4-glycosidic bonds of pectin polymers, and it has potential uses in the textile industry. In this study, a novel pectate lyase belonging to polysaccharide lyase family 10 was screened from the secreted enzyme extract of Paenibacillus polymyxa KF-1 and identified by liquid chromatography-MS/MS. The gene was cloned from P. polymyxa KF-1 genomic DNA and expressed in Escherichia coli. The recombinant enzyme PpPel10a had a predicted Mr of 45.2 kDa and pI of 9.41. Using polygalacturonic acid (PGA) as substrate, the optimal conditions for PpPel10a reaction were determined to be 50 °C and pH 9.0, respectively. The Km, vmax and kcat values of PpPel10a with PGA as substrate were 0.12 g/L, 289 μmol/min/mg, and 202.3 s−1, respectively. Recombinant PpPel10a degraded citrus pectin, producing unsaturated mono- and oligogalacturonic acids. PpPel10a reduced the viscosity of PGA, and weight loss of ramie (Boehmeria nivea) fibers was observed after treatment with the enzyme alone (22.5%) or the enzyme in combination with alkali (26.3%). This enzyme has potential for use in plant fiber processing.

A new cold-active and alkaline pectate lyase from Antarctic bacterium with high catalytic efficiency

2019 ◽  
Vol 103 (13) ◽  
pp. 5231-5241 ◽  
Author(s):  
Yumeng Tang ◽  
Pan Wu ◽  
Sijing Jiang ◽  
Jonathan Nimal Selvaraj ◽  
Shihui Yang ◽  
...  
Keyword(s):  
Pectate Lyase ◽  
Catalytic Efficiency ◽  
Antarctic Bacterium ◽  
Cold Active ◽  
Alkaline Pectate Lyase

scholarly journals Cloning and Sequencing of a High-alkaline Pectate Lyase Gene from an AlkaliphilicBacillusIsolate

1999 ◽  
Vol 63 (6) ◽  
pp. 998-1005 ◽  
Author(s):  
Yuji HATADA ◽  
Norihiko HIGAKI ◽  
Kazuhiro SAITO ◽  
Akinori OGAWA ◽  
Kazuhisa SAWADA ◽  
...  
Keyword(s):  
Pectate Lyase ◽  
Cloning And Sequencing ◽  
Alkaline Pectate Lyase

Highly alkaline pectate lyase Pel-4A from alkaliphilic Bacillus sp. strain P-4-N: its catalytic properties and deduced amino acid sequence

Extremophiles ◽  
2000 ◽  
Vol 4 (6) ◽  
pp. 377-383 ◽  
Author(s):  
Tohru Kobayashi ◽  
Yuji Hatada ◽  
Atsushi Suzumatsu ◽  
Katsuhisa Saeki ◽  
Yoshihiro Hakamada ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Catalytic Properties ◽  
Pectate Lyase ◽  
Bacillus Sp ◽  
Alkaliphilic Bacillus ◽  
Alkaline Pectate Lyase
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