Co-expression, purification and characterization of the lipase and foldase of Burkholderia contaminans LTEB11

Advances in Recombinant Lipases: Production, Engineering, Immobilization and Application in the Pharmaceutical Industry

“…The co-expression of the lipase gene with chaperones could be a reliable tool for obtaining the overproduction of recombinant lipases. Alnoch and collaborators [73], for example, co-expressed the lipase LipBC and its foldase LifBC genes from the Gram-negative bacterium Burkholderia contaminans LTEB11 ( Table 1). E. coli BL21 (DE3) strain was used as expression host and the best lipolytic activity was verified with the co-expression of the N-terminal truncated lipase gene with the full-length foldase gene.…”

“…This combination resulted in an activity of 127 U/mL against olive oil. Interestingly, no lipolytic activity was found when the lifBC gene was not expressed, showing that this foldase plays a key role in lipase conformation and function [73]. Although only the lipBC gene was fused to a His-tag, both proteins were purified in a single affinity chromatography step, which could be attractive from an industrial perspective.…”

“…The foldase LifBC must have formed a complex with the lipase LipBC, which was bound to the column, through a hydrophobic interaction between them. The complex formed, rLipBC, had maximum specific activity of 1426 U/mg towards tributyrin, and maintained a high conversion rate (>80%) of ethyl-oleate in n-hexane over five reaction cycles of 6 h at 45 • C, when it was immobilized on a hydrophobic support (Sepabeads™ FP-BU) [73]. G55Y, T52Y and AMS8 recombinant lipases…”

Genome sequencing of Burkholderia contaminans LTEB11 reveals a lipolytic arsenal of biotechnological interest

“…Indeed, we have produced LipBC by submerged fermentation and by solid-state fermentation and applied it in esterification and transesterification reactions for biodiesel synthesis [10][11][12] and resolution of racemates [13]. Recently, genes encoding lipase LipBC (lipA) and foldase LifBC (lipB) were identified and coexpressed in Escherichia coli, with a recombinant Lip-LifBC complex being purified and characterized [14]. However, little is known about the genome of this bacterium and whether it might have other biotechnological applications.…”

Estudos estruturais e funcionais de uma lipase de Beauveria bassiana imobilizada e expressa em Aspergillus nidulans: sensibilidade de linhagens celulares de glioblastoma aos hidrolisados de óleos brasileiros

“…Em razão do que foi discutido sobre a utilização de lipases no mercado, a expressão heteróloga apresenta-se como medida a aumentar a obtenção deste insumo industrial, além de auxiliar nos estudos bioquímicos dessas enzimas. Já são descritos trabalhos detalhando a expressão de lipases em Escherichia coli (KRUGENER et al, 2009;ALNOCH et al, 2018), Aspergillus oryzae (HOEGH et al, 1995) e Trichoderma reesei (QIN et al, 2012) o que tornou a produção de lipases independente das matrizes oleaginosas, comumente utilizadas para induzir a expressão desta enzima em cepas selvagens. Entretanto algumas dificuldades são pertinentes que limitam a utilização destes sistemas heterólogos em escala laboratorial: baixa atividade lipolítica nestas cepas, necessidade de substâncias indutoras com alto custo financeiro, síntese de lipases inativas ou que necessitam de uma etapa adicional para ativação.…”