Cloning, expression, purification, and characterization of lipase 3646 from thermophilic indigenous Cohnella sp. A01

Golaki, Bahram pooreydy; Aminzadeh, Saeed; Karkhane, Ali Asghar; Yakhchali, Bagher; Farrokh, Parisa; Khaleghinejad, Seyed Hossein; Tehrani, Asal Akhavian; Mehrpooyan, Sina

doi:10.1016/j.pep.2014.10.002

Cited by 34 publications

(21 citation statements)

References 37 publications

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“…The thermal stability of TLip was better than lipase 3646 from thermophilic bacterium Cohnella sp. A01, which exhibited about 58% of its activity after incubation at 50 °C for 2 H .…”

Section: Resultsmentioning

confidence: 99%

Identification and characterization of a novel esterase from Thauera sp.

Yang

Guo

et al. 2018

Biotech and App Biochem

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A novel esterase gene TLip was identified from the strain Thauera sp. and expressed at high levels in Escherichia coli. The TLip protein shared the highest identity (48%) to esterase TesA from Pseudomonas aeruginosa when compared to enzymes with reported properties. Phylogenetic analysis showed that TLip belongs to the GDSL family of bacterial lipolytic enzymes. TLip was an alkaline esterase with a broad optimal temperature range 37-50 °C and an optimal pH of 8.0. Substrate specificity assays showed that TLip preferred medium chain p-nitrophenyl esters (C -C ). Besides, the activity of TLip was strongly inhibited by Cu but greatly enhanced by Triton X-100 and Tween 80. Thermostability assay revealed that TLip was stable without loss of activity at 37 °C and still retained 69% activity at 50 °C after 2 H of incubation. Together, these provided a good candidate for further exploration of TLip as a promising biocatalyst in industry.

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“…The thermal stability of TLip was better than lipase 3646 from thermophilic bacterium Cohnella sp. A01, which exhibited about 58% of its activity after incubation at 50 °C for 2 H .…”

Section: Resultsmentioning

confidence: 99%

Identification and characterization of a novel esterase from Thauera sp.

Yang

Guo

et al. 2018

Biotech and App Biochem

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“…pullulan hydrolyzing enzyme was activated after CaCl 2 addition [28, 32–34]. Additionally, Ca 2+ and Mg 2+ improved the temperature stability of the enzyme [35, 36]. Some other cations, Li + , Na + , K + , Co 2+ , and Cu 2+ , severely reduced enzyme activity.…”

Section: Discussionmentioning

confidence: 99%

“…hydrothermalis did not affect by Na + and Mg 2+ while Mn 2+ increase its activity. Metal cations inhibitory effect such as Ni 2+ and Cu 2+ has been seen for almost all amylopullulanases [6, 18, 28, 31, 36, 37]. …”

Section: Discussionmentioning

confidence: 99%

Cohnella amylopullulanases: Biochemical characterization of two recombinant thermophilic enzymes

et al. 2017

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Some industries require newer, more efficient recombinant enzymes to accelerate their ongoing biochemical reactions in harsh environments with less replenishment. Thus, the search for native enzymes from extremophiles that are suitable for use under industrial conditions is a permanent challenge for R & D departments. Here and toward such discoveries, two sequences homologous to amylopullulanases (EC 3.2.1.41, GH57) from an endogenous Cohnella sp., [Coh00831 (KP335161; 1998 bp) and Coh01133 (KP335160: 3678 bp)] were identified. The genes were heterologously expressed in E. coli to both determine their type and further characterize their properties. The isolated DNA was PCR amplified with gene specific primers and cloned in pET28a, and the recombinant proteins were expressed in E. coli BL21 (DE3). The temperatures and pH optima of purified recombinants Coh 01133 and Coh 00831 enzymes were 70°C and 8, and 60°C and 6, respectively. These enzymes are stable more than 90% in 60°C and 50°C for 90 min respectively. The major reactions released sugars which could be fractionated by HPLC analysis, from soluble starch were mainly maltose (G2), maltotriose (G3) and maltotetraose (G4). The enzymes hydrolyzed pullulan to maltotriose (G3) only. Enzyme activities for both proteins were improved in the availability of Mn2+, Ba2+, Ca2+, and Mg2+ and reduced in the presence of Fe2+, Li2+, Na2+, Triton X100 and urea. Moreover, Co2+, K+, and Cu2+ had a negative effect only on Coh 01133 enzyme.

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“…Golaki et al. reported 60% of residual activity at 60 °C using a lipase from Thermophilic indigenous Cohnella sp. A01.…”

Section: Discussionmentioning

confidence: 99%

Production and application of a thermostable lipase from Serratia marcescens in detergent formulation and biodiesel production

García‐Silvera

Martínez‐Morales

Bertrand

et al. 2017

Biotech and App Biochem

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In this study, extracellular lipase was produced by Serratia marcescens wild type and three mutant strains. The maximum lipase activity (80 U/mL) was obtained with the SMRG4 mutant strain using soybean oil. Using a 2 factorial design, the lipase production increased 1.55-fold (124 U/mL) with 4% and 0.05% of soybean oil and Triton X-100, respectively. The optimum conditions for maximum lipase activity were 50 °C and pH 8. However, the enzyme was active in a broad range of pH (6-10) and temperatures (5-55 °C). This lipase was stable in organic solvents and in the presence of oxidizing agents. The enzyme also proved to be efficient for the removal of triacylglycerol from olive oil in cotton cloth. A Box-Behnken experimental design was used to evaluate the effects of the interactions between total lipase activity, buffer pH, and wash temperatures on oil removal. The model obtained suggested that all selected factors had a significant impact on oil removal, with optimum conditions of 550 U lipase, 45 °C, pH 9.5, with 79.45% removal. Biotransformation of waste frying oil using the enzyme and in presence of methanol resulted in the synthesis of methyl esters such as methyl oleate, methyl palmitate, and methyl stearate.

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Cloning, expression, purification, and characterization of lipase 3646 from thermophilic indigenous Cohnella sp. A01

Cited by 34 publications

References 37 publications

Identification and characterization of a novel esterase from Thauera sp.

Identification and characterization of a novel esterase from Thauera sp.

Cohnella amylopullulanases: Biochemical characterization of two recombinant thermophilic enzymes

Production and application of a thermostable lipase from Serratia marcescens in detergent formulation and biodiesel production

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