Thermodynamic characterization of a highly thermoactive extracellular pectate lyase from a new isolate Bacillus pumilus DKS1

Basu, Snehasish; Ghosh, Abhrajyoti; Bera, Amit; Saha, Manabendra N.; Chattopadhyay, Dhrubajyoti; Chakrabarti, Krishanu

doi:10.1016/j.biortech.2008.03.032

Cited by 33 publications

(31 citation statements)

References 33 publications

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“…The maximum activity of pectate lyase (25 U/ml) in the medium was observed after 24 h of growth in the presence of ramie Wbres ( Table 1). The DKS1 pectate lyase has been puriWed and also characterised [12]. Neither pectin esterase (EC 3.1.1.11) nor polygalacturonase (EC 3.2.1.15) was detected in the cell-free culture Xuid.…”

Section: Resultsmentioning

confidence: 99%

“…The pectinase enzyme selectively degraded only the noncellulosic gummy material of Wbres causing 10.96% Wbre weight loss after 24 h. This weight loss was lower than that obtained using Bacillus pumilus DKS1. However, 0.04% NaOH was used during the degumming by using Bacillus pumilus dcsr1, which was lower than the alkali used in degumming made by Bacillus pumilus DKS1 [12]. Again according to the procedure of Zheng et al [19] using alkalophilic Bacillus sp.…”

Section: Resultsmentioning

confidence: 99%

“…The samples were analysed for pectate lyase activity by the method described by Basu et al [12]. Detectable enzyme activity was observed only after 72 h. Thereafter enzyme activity increased almost seven times up to 10 days (Fig.…”

Section: Pectate Lyase Activity In Large-scale Degummingmentioning

confidence: 99%

“…At the laboratory scale the salt-papaya medium was sterilised by a standard autoclaved method (at 121°C or 249°F for 15 min). The Xask was incubated at 30°C on a rotary shaker (150 rev/min) for 24 h. The culture was centrifuged (7,000 rpm, 10 min, 4°C) and the cell-free supernatant was assayed for pectate lyase activity by the TBA method (A 550 ) according Basu et al [12] based on earlier methods [13,14]. A control system was run without adding the microbial culture.…”

Section: Culture Conditions For Degumming On Small Scalementioning

confidence: 99%

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Large-scale degumming of ramie fibre using a newly isolated Bacillus pumilus DKS1 with high pectate lyase activity

Basu¹,

Saha²,

Chattopadhyay³

et al. 2008

J Ind Microbiol Biotechnol

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A combined (enzymatic and chemical) process using a Bacillus pumilus strain (DKS1), isolated from the soil, was used to degum ramie bast fibres. After 24 h of incubation with the isolated pectinolytic strain using a low-cost medium, the weight loss of the ramie fibre was found to be 25% under small scale. High activity of pectate lyase was detected in the culture supernatants; 400 kg of ramie fibres was degummed with 24% weight loss in large-scale degumming under field conditions. No cellulase activity was found. Microbial intervention followed by mild (0.1%) alkali treatment showed high percentage of weight loss from the ramie fibre. Bacterial degumming followed by chemical treatment resulted in an increase of single fibre tenacity (cN/tex) by more than 20.81% as compared to non-degummed (decorticated) fibre samples. Scanning electron micrographs (SEM) and fluorescence microscope showed that after Bacillus pumilus DKS1 treatment the surface of the decorticated ramie fibre becomes very smooth. These results indicate the process provides an economical and eco-friendly method for the small scale as well as large-scale degumming of decorticated ramie fibre. This study has great relevance to the textile as well as paper industry.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Pectate Lyase Activity In Large-scale Degummingmentioning

confidence: 99%

Section: Culture Conditions For Degumming On Small Scalementioning

confidence: 99%

See 2 more Smart Citations

Large-scale degumming of ramie fibre using a newly isolated Bacillus pumilus DKS1 with high pectate lyase activity

Basu¹,

Saha²,

Chattopadhyay³

et al. 2008

J Ind Microbiol Biotechnol

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“…In previous papers (Basu et al 2008(Basu et al , 2009a we had reported the isolation and characterization of a theromoactive pectate lyase from a strain of Bacillus pumilus DKS1 and its potential use for degumming ramie fiber. This paper reports the cloning, expression, sequencing and site-directed mutagenesis of pel gene encoding pectate lyase from Bacillus pumilus DKS1 strain (Basu et al 2008) to elucidate that Arg235 is an essential catalytic residue to degumming ramie fibre.…”

Section: Introductionmentioning

confidence: 99%

Arg235 is an essential catalytic residue of Bacillus pumilus DKS1 pectate lyase to degum ramie fibre

et al. 2010

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After 24 h of incubation with only purified pectate lyase isolated from Bacillus pumilus DKS1 (EF467045), the weight loss of the ramie fibre was found to be 25%. To know the catalytic residue of pectate lyase the pel gene encoding a pectate lyase from the strain Bacillus pumilus DKS1 was cloned in E. coli XL1Blue and expressed in E. coli BL21 (DE3) pLysS. The pel gene was sequenced and showed 1032 bp length. After purification using CM-Sepharose the enzyme showed molecular weight of 35 kDa and maximal enzymatic activity was observed at 60°C and a pH range of 8.5-9.0. Both Ca²(+) and Mn²(+) ions were required for activity on Na-pectate salt substrates, while the enzyme was strongly inhibited by Zn²(+) and EDTA. The deduced nucleotide sequence of the DKS1 pectate lyase (EU652988) showed 90% homology to pectate lyases from Bacillus pumilus SAFR-032 (CP000813). The 3D structure as well as the catalytic residues was predicted using EasyPred software and Catalytic Site Atlas (CSA), respectively. Site directed mutagenesis confirmed that arginine is an essential catalytic residue of DKS1 pectate lyase.

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Biotechnological Applications of Biocatalysts from the Firmicutes Bacillus and Geobacillus Species

Satyanarayana

Sharma

Mehta

et al. 2011

Microorganisms in Sustainable Agriculture and Biotechnology

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Thermodynamic characterization of a highly thermoactive extracellular pectate lyase from a new isolate Bacillus pumilus DKS1

Cited by 33 publications

References 33 publications

Large-scale degumming of ramie fibre using a newly isolated Bacillus pumilus DKS1 with high pectate lyase activity

Large-scale degumming of ramie fibre using a newly isolated Bacillus pumilus DKS1 with high pectate lyase activity

Arg235 is an essential catalytic residue of Bacillus pumilus DKS1 pectate lyase to degum ramie fibre

Biotechnological Applications of Biocatalysts from the Firmicutes Bacillus and Geobacillus Species

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