Characteristics of Polygalacturonate Lyase C from Bacillus subtilis 7-3-3 and Its Synergistic Action with PelA in Enzymatic Degumming

Zou, Mouyong; Li, Xuezhi; Zhao, Jian; Qu, Yinbo

doi:10.1371/journal.pone.0079357

Cited by 21 publications

(18 citation statements)

References 17 publications

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“…The degumming efficiency of M3 was among the highest of the alkaline and thermostable pectate lyases hitherto reported (Basu et al 2009;Brühlmann et al 2000;Kapoor et al 2001;Zou et al 2013a). Moreover, the ramie degumming efficiency could be further improved by using M3 mutant in conjunction with other enzymes, such as xylanase or other pectate lyases (Zou et al 2013b). Overall, our findings indicate that the M3 mutant enzyme can be efficiently used for degumming ramie via the combined enzyme-chemical process, confirming its great potentials in applications in textile industries.…”

Section: Discussionmentioning

confidence: 95%

“…For example, alkaline Pels have important industrial applications in textile and food production industries and are used for degumming ramie. As a potential alternative to chemical degumming process, the advantages of enzymatic degumming include flexible and mild processing conditions, limited damage to fibers, environmentally friendly operation, and easy quality control (Zou et al 2013b). However, thus far, enzymatic degumming in the textile industry has been hampered by the high cost and partial degumming performance of the currently available enzymes.…”

Section: Introductionmentioning

confidence: 99%

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

Liang

Gui

Zhou

et al. 2014

Appl Microbiol Biotechnol

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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 13 h. In contrast, the wild-type enzyme was completely inactivated in less than 10 min 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.

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Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

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

Liang

Gui

Zhou

et al. 2014

Appl Microbiol Biotechnol

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“…Thus, the use of pectinase activity alone is significantly inadequate to screen bacteria for ramie degumming, and the practical application effects of ramie degumming should be combined. 29 As a primary key factor of ''primase'' in biological degumming, pectinase directly influences the ramiedegumming effect. 30 In this study, the pectinase activities of the two dominant strains (2-1 and 4-1) were 85.1 and 98.2 U/mg at 20 h, and the residual gum contents of ramie were 8.32% and 10.86% at 14 h, respectively.…”

Section: Discussionmentioning

confidence: 99%

Screening and identification of pectinolytic bacteria for ramie degumming

Cheng

Duan

Feng

et al. 2020

Textile Research Journal

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To explore high-quality microbial resources with the capability of ramie degumming, we collected soil samples from rotten ramie and straw heaps. After enrichment culture by ramie raw materials, bacterial strains with the potential ramie-degumming function were screened using a pectin-hydrolysis plate. Dominant bacteria were identified by combining colonial morphological characteristics with the molecular biology method, and their ramie-degumming effects were verified through comprehensive biological degumming indices. Results demonstrated that Bacillus aryabhattai, Bacillus thuringiensis, Lysinibacillus fusiformis, and Acidovorax temperans were successfully obtained. The highest pectinase activity, 98.2 U/mg, was found by A. temperans. B. thuringiensis showed the best ramie-degumming effect. The residual gum content, single-fiber linear density, and bundle-breaking strength of the degummed ramie fiber treated with B. thuringiensis were 8.32%, 6.80 dtex, and 7.84 cN/dtex, respectively. The residual gum content of the ramie fiber treated with B. thuringiensis met the textile requirement (<10%), and the values of all other indicators were also satisfactory. Therefore, B. thuringiensis was an excellent strain for ramie degumming, indicating potential industrial applications.

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“…However, the total gum content of ramie fibers used in this experiment was ϳ34%, and only fibers with Ͻ2.5% residual gum, which means those with at least Ͼ30% weight loss, can be directly used in further textile processes (53). The enzyme-chemical degumming efficiency of mutant EA was close to the efficiency (at least Ͼ30% weight loss) of the traditional chemical degumming method which had been widely used in the ramie textile industry, and moreover, the ramie degumming efficiency could be further improved by using mutant EA in conjunction with other enzymes, such as xylanase or other Pels (23), and subsequent processes, such as bleaching, in the future.…”

Section: Discussionmentioning

confidence: 83%

“…Alkaline Pels in particular are being introduced in textile industrial processing and degumming to facilitate the release of fibers from flax, ramie, sunn hemp, buel, and jute (15)(16)(17)(18)(19) as an alternative to conventional retting. Compared with conventional chemical degumming processes, enzymatic degumming has several advantages, including flexible and mild processing conditions, limited damage to fibers, environmentally friendly operation, and easy quality control (20)(21)(22)(23). However, enzymatic degumming has not been widely used in the textile industry because of the high cost and unsatisfactory degumming efficiency of currently available enzymes.…”

mentioning

confidence: 99%

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

Zhou

Xue

et al. 2015

Appl Environ Microbiol

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c Thermostable 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 from Bacillus sp. 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 Ca 2؉ . 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 the t 50 (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 the t 50 value 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. P ectin, a naturally ubiquitous constituent of the middle lamella of the primary cell wall in plants, is a heteropolysaccharide composed of ␣-1,4-linked galacturonate chains with a high percentage of methyl esterification and functions to form a network with other noncellulosic materials, such as proteins and waxes (1-3). Pectin degradation requires the combined action of several enzymes that can be classified into two main groups: methylesterases, which remove methoxyl groups from pectin, and depolymerases (hydrolases and lyases), which cleave the bonds between galacturonate units (4-6).Pectate lyases (Pels) (EC 4.2.2.2) cleave ␣-1,4-linked galacturonate units of pectate by ␤-elimination, giving rise to an unsaturated C-4 -C-5 bond at the nonreducing end of the newly formed oligogalacturonate (7). Pels are widely distributed among microbial plant pathogens and have been the focus of several studies aiming to elucidate the roles of these enzymes as virulence factors (8, 9). Pels have also been found in some other microorganisms, including bacteria of the genus Bacillus (10) and some thermophilic bacteria (11). In general, Pels work efficiently at alkaline pH (between pH 8 and 10), and additional Ca 2ϩ ions are required for efficient enz...

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Characteristics of Polygalacturonate Lyase C from Bacillus subtilis 7-3-3 and Its Synergistic Action with PelA in Enzymatic Degumming

Cited by 21 publications

References 17 publications

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

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

Screening and identification of pectinolytic bacteria for ramie degumming

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

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