Production of alkaline pectinase: a case study investigating the use of tobacco stalk with the newly isolated strain Bacillus tequilensis CAS-MEI-2-33

Zhang, Ge; Li, Shugui; Wang, Juan; Wang, Fan; Xin, Yu‐Hua; Shen, Zhong Yao; Zhang, Haibo; Ma, Ming; Liu, Haobao

doi:10.1186/s12896-019-0526-6

Cited by 16 publications

(4 citation statements)

References 30 publications

(40 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Pectin lyase is a highly valuable industrial enzyme that has been widely used in various important fields, such as fruit juice processing and extraction, cotton and linen textile dyeing and processing, plant fiber degumming, environmental industrial wastewater treatment, and pulp and paper production (Bruhlmann et al, 2000;Phugare et al, 2011;Zhang et al, 2019;Jaworska et al, 2022). As more attention is given to the environment, reducing pollution and energy consumption has become an urgent issue in traditional industries.…”

Section: Introductionmentioning

confidence: 99%

Flow-cytometric cell sorting coupled with UV mutagenesis for improving pectin lyase expression

Fang,

Ma,

Wang

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Introduction: Alkaline pectin lyase is an important enzyme with a wide range of applications in industrial production, It has been widely used in many important fields such as fruit juice processing and extraction, the dyeing and processing of cotton and linen textiles, degumming plant fibers, environmental industrial wastewater treatment, and pulp and paper production. PGLA-rep4 was previously generated as a modified alkaline pectin lyase with high specific activity at pH 11.0°C and 70°C. However, the pre-constructed high-activity pectin lyase expression strains are still difficult to apply in industrial production due to their limited enzymatic activity. We hope to solve these problems by combining modern breeding techniques with high-throughput equipment to rapidly screen alkaline pectin lyase with higher enzymatic activity and lower cost.Methods: We fused the genes encoding PGLA-rep4 and fluorescent protein egfp using a flexible linker peptide and ligated them into a temperature-sensitive plasmid, pKD46. The constructed screening plasmid pKD46-PGLA-rep4-egfp was then transformed into an expression host and screened via flow-cytometric cell sorting coupled with UV mutagenesis.Results: Following mutagenesis, primary screening, and secondary screening, the high-expression strain, named Escherichia coli BL21/1G3, was obtained. The screening plasmid pKD46-PGLA-rep4-egfp was eliminated, and the original expression plasmid pET28a-PGLA-rep4 was then retransformed into the mutant strains. After induction and fermentation, pectin lyase activity in E. coli BL21/1G3 was significantly increased (1.37-fold relative to that in the parental E. coli BL21/PGLA-rep4 strain, p < 0.001), and the highest activity was 230, 240 U/mL at 144 h. Genome sequencing revealed that genes encoding ribonuclease E (RNase E) and diadenosine tetraphosphatase (ApaH) of E. coli BL21/1G3 were mutated compared to the sequence in the original E. coli BL21 (DE3) strain, which could be associated with increased enzyme expression.Discussion: Our work provides an effective method for the construction of strains expressing pectin lyase at high levels.

show abstract

Section: Introductionmentioning

confidence: 99%

Flow-cytometric cell sorting coupled with UV mutagenesis for improving pectin lyase expression

Fang,

Ma,

Wang

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…If applied to ramie bio-degumming, alkaline pectate lyase will yield high efficiency, low cost and low pollution. Moreover, it can reserve the inherent morphology of ramie fibers and improve their quality [ 8 ]. In addition, research findings show that Ca 2+ can bind to pectate lyase and substrates to promote the enzyme-substrate reaction, so as to improve the activity of pectate lyase [ 9 , 10 ].…”

Section: Introductionmentioning

confidence: 99%

Improvement on Thermostability of Pectate Lyase and Its Potential Application to Ramie Degumming

Feng

Yang

et al. 2022

Polymers

View full text Add to dashboard Cite

In order to obtain a thermostable pectate lyase for ramie degumming, a rational design based on structural analysis was carried out on a novel pectate lyase (Pel419) derived from the Dickea dadantii DCE-01 for high-efficiency ramie degumming. A total of five potential amino acid sites were chosen to replace residues. Then, the mutant enzymes were subjected to the heterologous expressions in Escherichia coli and their enzymatic characteristics were determined. The optimal reaction temperature for the five mutants kept consistent with that for the wild type. The enzyme activity and thermal stability of mutant V52A were significantly improved. Meanwhile, the weight loss rate obtained by V52A with the best enzymatic characteristics in the ramie degumming process at 50 °C is comparable with that obtained by commercial cotton-ramie processing pectinases, indicating that V52A was a potential industrial enzyme that could be applied to large-scale ramie degumming. In this study, the biological functions of conservative residues of Pel419 were preliminarily explored. The mutant V52A with both enzymatic activity and improved heat resistance was acquired, providing a superior material for developing enzyme preparations of ramie degumming, and rendering an effective method for the rational design aiming to improve the thermostability of pectate lyase.

show abstract

“…Therefore, most of them are left to decay in the fields or are burnt [3]. Treatment of agricultural wastes is not only a concern for the environment but it also affects the development of sustainable agriculture [4][5][6].…”

Section: Introductionmentioning

confidence: 99%

“…Exploring microorganisms and enzymes associated with the degradation of biomass by traditional methods or functional metagenomic approaches can have great potentials for sustainable agriculture and biofuel production [4,[6][7][8]. These vegetable wastes are made-up of cellulose, hemicellulose and lignin.…”

Section: Introductionmentioning

confidence: 99%

Genomic, Transcriptomic and Enzymatic Insight into Lignocellulolytic System of a Plant Pathogen Dickeya sp. WS52 to Digest Sweet Pepper and Tomato Stalk

et al. 2019

View full text Add to dashboard Cite

Dickeya sp., a plant pathogen, causing soft rot with strong pectin degradation capacity was taken for the comprehensive analysis of its corresponding biomass degradative system, which has not been analyzed yet. Whole genome sequence analysis of the isolated soft-rotten plant pathogen Dickeya sp. WS52, revealed various coding genes which involved in vegetable stalk degradation-related properties. A total of 122 genes were found to be encoded for putative carbohydrate-active enzymes (CAZy) in Dickeya sp. WS52. The number of pectin degradation-related genes, was higher than that of cellulolytic bacteria as well as other Dickeya spp. strains. The CAZy in Dickeya sp.WS52 contains a complete repertoire of enzymes required for hemicellulose degradation, especially pectinases. In addition, WS52 strain possessed plenty of genes encoding potential ligninolytic relevant enzymes, such as multicopper oxidase, catalase/hydroperoxidase, glutathione S-transferase, and quinone oxidoreductase. Transcriptome analysis revealed that parts of genes encoding lignocellulolytic enzymes were significantly upregulated in the presence of minimal salt medium with vegetable stalks. However, most of the genes were related to lignocellulolytic enzymes, especially pectate lyases and were downregulated due to the slow growth and downregulated secretion systems. The assay of lignocellulolytic enzymes including CMCase and pectinase activities were identified to be more active in vegetable stalk relative to MSM + glucose. However, compared with nutrient LB medium, it needed sufficient nutrient to promote growth and to improve the secretion system. Further identification of enzyme activities of Dickeya sp.WS52 by HPLC confirmed that monosaccharides were produced during degradation of tomato stalk. This identified degradative system is valuable for the application in the lignocellulosic bioenergy industry and animal production.

show abstract

Production of alkaline pectinase: a case study investigating the use of tobacco stalk with the newly isolated strain Bacillus tequilensis CAS-MEI-2-33

Cited by 16 publications

References 30 publications

Flow-cytometric cell sorting coupled with UV mutagenesis for improving pectin lyase expression

Flow-cytometric cell sorting coupled with UV mutagenesis for improving pectin lyase expression

Improvement on Thermostability of Pectate Lyase and Its Potential Application to Ramie Degumming

Genomic, Transcriptomic and Enzymatic Insight into Lignocellulolytic System of a Plant Pathogen Dickeya sp. WS52 to Digest Sweet Pepper and Tomato Stalk

Contact Info

Product

Resources

About