Cell-bound lipases from Burkholderia sp. ZYB002: gene sequence analysis, expression, enzymatic characterization, and 3D structural model

Abstract

BackgroundThe whole-cell lipase from Burkholderia cepacia has been used as a biocatalyst in organic synthesis. However, there is no report in the literature on the component or the gene sequence of the cell-bound lipase from this species. Qualitative analysis of the cell-bound lipase would help to illuminate the regulation mechanism of gene expression and further improve the yield of the cell-bound lipase by gene engineering.ResultsThree predictive cell-bound lipases, lipA, lipC21 and lipC24, from Burkholderia sp. ZYB002 were cloned and expressed in E. coli. Both LipA and LipC24 displayed the lipase activity. LipC24 was a novel mesophilic enzyme and displayed preference for medium-chain-length acyl groups (C10-C14). The 3D structural model of LipC24 revealed the open Y-type active site. LipA displayed 96 % amino acid sequence identity with the known extracellular lipase. lipA-inactivation and lipC24-inactivation decreased the total cell-bound lipase activity of Burkholderia sp. ZYB002 by 42 % and 14 %, respectively.ConclusionsThe cell-bound lipase activity from Burkholderia sp. ZYB002 originated from a multi-enzyme mixture with LipA as the main component. LipC24 was a novel lipase and displayed different enzymatic characteristics and structural model with LipA. Besides LipA and LipC24, other type of the cell-bound lipases (or esterases) should exist.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-016-0269-6) contains supplementary material, which is available to authorized users.

Purification and characterization of alkaline lipase production byPseudomonas aeruginosaHFE733 and application for biodegradation in food wastewater treatment

“…The lipase exhibited the highest activity at 40 C, which was similar to the corresponding enzymes from Burkholderia sp. ZYB002 [24] and P. aeruginosa LX1 [5]. The purified lipase was stable between 30 and 40 C, retaining approximately 70% of its activity, but the thermal stability of the enzyme declined at 50-70 C (Figure 6(b)).…”

High-level soluble expression and enzymatic characterization of Burkholderia sp. lipase with steryl ester hydrolysis activity in E. coli

“…The amino acid sequence identity was over 93% between steryl esterase from B. cepacia ST200 and LipA from Burkholderia sp. ZYB002 [49,50]. In this research, both lipase activity and steryl esterase activity were determined from LipA from Burkholderia sp.…”

“…Recombinant plasmids pETDuet-A1B2, pACYCDuet-A1, and pETDuet-B2 were derived from pEDSF-lipB-lipA, respectively [50]. Plasmid pEDSF-lipB-lipA was firstly double digested by the restriction endonucleases, BamH Ⅰ and Hind Ⅲ, followed by Bgl Ⅱ and Xho Ⅰ digestion.…”

“…Table 3 were constructed as follows. The DNA fragments of lipA and lipB were amplified by PCR using plasmid pMD18T-lipAB as the template [50]. The oligonucleotide sequences of PCR primers, PCR primer pairs, annealing temperatures, and PCR products were listed in Table 4.…”

Heterologous Expression and Biochemical Characterization of Lipase from Burkholderia cepacia Lu10-1

“…It has been reported that the lipase from Burkholderia (previously Pseudomonas) cepacia possesses attractive particular interest due to its high thermostability and strong tolerance to organic solvents (Jia et al, 2010; Mello Bueno et al, 2015;Sasso et al, 2016), therefore, different kinds of Burkholderia lipase genes have been cloned and over-expressed in heterogonous or homologous hosts (Jia et al, 2010;Shu et al, 2016;Xie et al, 2016;Yang et al, 2007). However, the high yield of the active forms in heterogonous host such as Escherichia coli has not yet been achieved.…”