Cloning, expression and characterization of a metagenome derived thermoactive/thermostable pectinase

Singh, Rahul Kumar; Dhawan, Samriti; Singh, Kashmir; Kaur, Jagdeep

doi:10.1007/s11033-012-1685-x

Cited by 44 publications

(19 citation statements)

References 57 publications

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“…The three groups are characterized by having different catalytic types (1st group: exo -PGases (EPG; EC 3.2.1.67); 2nd group: exo -PGases (EPGD; EC 3.2.1.82); 3rd: endo -PGases (PG; EC 3.2.1.15)). Our results suggest that ACM61449 and ACM60667 belong to the 1st group, and may exhibit EPG activity, similar to what has been reported for AAD35522 (PelB) [18] or ACL36472 (PecJKR01) [29]. …”

Section: Resultssupporting

confidence: 87%

“…CbPelA showed higher thermal-stability than most reported EPGs (Table 1). The half-life of PecJKR01 at 60 °C is 5 h [29]. The bacterial Pgu B [8] retained about 20% of its activity at 60 °C for less than 10 min.…”

Section: Resultsmentioning

confidence: 99%

“…For example, Pgu B is completely inhibited by Cu 2+ [8], while PG is able to retain 50% of its activity [13]. Moreover, Mg 2+ or Co 2+ enhances the activity of PG, while Ca 2+ has no significant effect on both PG and PecJKR01 [13,29]. Therefore, the interaction between the EPGs and divalent metal cations is complex, and further investigation is needed to clarify the mechanism.…”

Section: Resultsmentioning

confidence: 99%

“…TS 47 [26]. However, only two thermophilic EPGs have been isolated from thermophilies [27–29]. One of the thermo-active EPGs that was identified was PelB cloned from T. maritime.…”

Section: Introductionmentioning

confidence: 99%

“…Another thermophilic pectinase, PecJKR01, was isolated from a soil metagenome sample. PecJKR01 demonstrated activity over a broad range of temperature and pH [29]. …”

Section: Introductionmentioning

confidence: 99%

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Cloning, Expression and Characterization of a Novel Thermophilic Polygalacturonase from Caldicellulosiruptor bescii DSM 6725

Chen

Sun

Zhou

et al. 2014

IJMS

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We cloned the gene ACM61449 from anaerobic, thermophilic Caldicellulosiruptor bescii, and expressed it in Escherichia coli origami (DE3). After purification through thermal treatment and Ni-NTA agarose column extraction, we characterized the properties of the recombinant protein (CbPelA). The optimal temperature and pH of the protein were 72 °C and 5.2, respectively. CbPelA demonstrated high thermal-stability, with a half-life of 14 h at 70 °C. CbPelA also showed very high activity for polygalacturonic acid (PGA), and released monogalacturonic acid as its sole product. The Vmax and Km of CbPelA were 384.6 U·mg−1 and 0.31 mg·mL−1, respectively. CbPelA was also able to hydrolyze methylated pectin (48% and 10% relative activity on 20%–34% and 85% methylated pectin, respectively). The high thermo-activity and methylated pectin hydrolization activity of CbPelA suggest that it has potential applications in the food and textile industry.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…TS 47 [26]. However, only two thermophilic EPGs have been isolated from thermophilies [27–29]. One of the thermo-active EPGs that was identified was PelB cloned from T. maritime.…”

Section: Introductionmentioning

confidence: 99%

“…Another thermophilic pectinase, PecJKR01, was isolated from a soil metagenome sample. PecJKR01 demonstrated activity over a broad range of temperature and pH [29]. …”

Section: Introductionmentioning

confidence: 99%

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