N-linked glycosylation of thermostable lipase from Bacillus thermocatenulatus to improve organic solvent stability

Kajiwara, Shota; Yamada, R.; Matsumoto, Takuya; Ogino, Hiroyasu

doi:10.1016/j.enzmictec.2019.109416

Cited by 16 publications

(3 citation statements)

References 32 publications

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“…Lipase is a hydrolase which acts on the ester bond with catalytic diversity, and it is also one of the most widely used biocatalysts in industrial production (Melani et al 2020 ). Geobacillus thermocatenulatus lipase 2 (GTL2) is a typical thermophilic enzyme with high catalytic activity and good thermal stability and has several applications in pharmaceutical and organic synthesis, chiral compound resolution, and bioenergy; thus, it is a promising biocatalyst with great application potential (Godoy et al 2019 ; Kajiwara et al 2020 ). In addition, Escherichia coli has a short culture cycle with high target protein levels, which makes it a good host for expressing GTL2.…”

Section: Introductionmentioning

confidence: 99%

A novel strategy for D-psicose and lipase co-production using a co-culture system of engineered Bacillus subtilis and Escherichia coli and bioprocess analysis using metabolomics

Luo

Wang

Chen

et al. 2021

Bioresour. Bioprocess.

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To develop an economically feasible fermentation process, this study designed a novel bioprocess based on the co-culture of engineered Bacillus subtilis and Escherichia coli for the co-production of extracellular D-psicose and intracellular lipase. After optimizing the co-culture bioprocess, 11.70 g/L of D-psicose along with 16.03 U/mg of lipase was obtained; the glucose and fructose were completely utilized. Hence, the conversion rate of D-psicose reached 69.54%. Compared with mono-culture, lipase activity increased by 58.24%, and D-psicose production increased by 7.08%. In addition, the co-culture bioprocess was explored through metabolomics analysis, which included 168 carboxylic acids and derivatives, 70 organooxygen compounds, 34 diazines, 32 pyridines and derivatives, 30 benzene and substituted derivatives, and other compounds. It also could be found that the relative abundance of differential metabolites in the co-culture system was significantly higher than that in the mono-culture system. Pathway analysis revealed that, tryptophan metabolism and β-alanine metabolism had the highest correlation and played an important role in the co-culture system; among them, tryptophan metabolism regulates protein synthesis and β-alanine metabolism, which is related to the formation of metabolic by-products. These results confirm that the co-cultivation of B. subtilis and E. coli can provide a novel idea for D-psicose and lipase biorefinery, and are beneficial for the discovery of valuable secondary metabolites such as turanose and morusin.

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Section: Introductionmentioning

confidence: 99%

A novel strategy for D-psicose and lipase co-production using a co-culture system of engineered Bacillus subtilis and Escherichia coli and bioprocess analysis using metabolomics

Luo

Wang

Chen

et al. 2021

Bioresour. Bioprocess.

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“…Lipase 2 from Geobacillus thermocatenulatus (BTL2) belongs to the lipase family 1.5, which is a typical thermophilic lipase with excellent catalytic properties such as strong thermal stability, tolerance to organic solvents, and high substrate selectivity. Due to the interesting performance of lipase BTL2, in recent years, studies have mainly focused on heterologous expression (Yamada et al 2016), the relationship between structure and function (Kajiwara et al 2020), and improvement of enzyme activity (Godoy et al 2019). Therefore, there is an urgent need to construct an engineered bacterial platform that can efficiently express lipase for in-depth applicable exploration.…”

Section: Introductionmentioning

confidence: 99%

High-efficiency expression of the thermophilic lipase from Geobacillus thermocatenulatus in Escherichia coli and its application in the enzymatic hydrolysis of rapeseed oil

Xu¹,

Tian²,

et al. 2020

3 Biotech

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Long-chain fatty acids are widely used in food and chemical industries, and the enzymatic preparation of fatty acids is considered an environmentally friendly process. In the present study, long-chain fatty acids were prepared by the enzymatic hydrolysis of rapeseed oil with a genetically engineered lipase. Because thermophilic lipase has strong stability at higher temperatures, it was more suitable for the industrial production of long-chain fatty acids. Therefore, the thermophilic lipase BTL2 from Geobacillus thermocatenulatus was efficiently expressed in E. coli BL21(DE3) cells with an enzyme activity of 39.50 U/mg followed by gene codon optimisation. Experimental results showed that the recombinant lipase BTL2 exhibited excellent resistance to certain organic solvents (n-hexane, benzene, ethanol, and butanol). The metal cation Ca 2+ and the non-ionic surfactant Triton-100X enhanced enzyme activity by 7.36% and 56.21% respectively. Moreover, the acid value of the liberated long-chain fatty acids by hydrolysing rapeseed oil was approximately 161.64 mg KOH/g at 50 °C in 24 h, the hydrolytic conversion rate was 91.45%, and the productivity was approximately 6.735 mg KOH/g h. These results suggested that the recombinant lipase BTL2 has excellent hydrolytic performance for rapeseed oil and showed great potential for the enzymatic preparation of long-chain fatty acids.

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“…Later, this enzyme-immobilized on different supports via diverse methodologies-has been extensively tested on different substrates, some of them chiral [45][46][47][48][49][50][51][52][53][54][55][56][57][58][59][60][61][62][63][64], generally with moderate results. Additionally, chemical [55,58,[65][66][67] and genetic [56,[68][69][70][71] modifications of BTL2 lipase for improving its catalytic behavior (typically, to reduce the steric hindrance around the active site) have been also reported. Finally, different papers in recent literature have employed the reported 3D structure of BTL2 for performing molecular simulations aiming to rationalize its catalytic performance and stability [38,[72][73][74].…”

Section: Introductionmentioning

confidence: 99%

Biocatalysis at Extreme Temperatures: Enantioselective Synthesis of both Enantiomers of Mandelic Acid by Transesterification Catalyzed by a Thermophilic Lipase in Ionic Liquids at 120 °C

Ramos-Martín¹,

Khiari²,

Alcántara³

et al. 2020

Catalysts

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The use of biocatalysts in organic chemistry for catalyzing chemo-, regio- and stereoselective transformations has become an usual tool in the last years, both at lab and industrial scale. This is not only because of their exquisite precision, but also due to the inherent increase in the process sustainability. Nevertheless, most of the interesting industrial reactions involve water-insoluble substrates, so the use of (generally not green) organic solvents is generally required. Although lipases are capable of maintaining their catalytic precision working in those solvents, reactions are usually very slow and consequently not very appropriate for industrial purposes. Increasing reaction temperature would accelerate the reaction rate, but this should require the use of lipases from thermophiles, which tend to be more enantioselective at lower temperatures, as they are more rigid than those from mesophiles. Therefore, the ideal scenario would require a thermophilic lipase capable of retaining high enantioselectivity at high temperatures. In this paper, we describe the use of lipase from Geobacillus thermocatenolatus as catalyst in the ethanolysis of racemic 2-(butyryloxy)-2-phenylacetic to furnish both enantiomers of mandelic acid, an useful intermediate in the synthesis of many drugs and active products. The catalytic performance at high temperature in a conventional organic solvent (isooctane) and four imidazolium-based ionic liquids was assessed. The best results were obtained using 1-ethyl-3-methyl imidazolium tetrafluoroborate (EMIMBF4) and 1-ethyl-3-methyl imidazolium hexafluorophosphate (EMIMPF6) at temperatures as high as 120 °C, observing in both cases very fast and enantioselective kinetic resolutions, respectively leading exclusively to the (S) or to the (R)-enantiomer of mandelic acid, depending on the anion component of the ionic liquid.

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N-linked glycosylation of thermostable lipase from Bacillus thermocatenulatus to improve organic solvent stability

Cited by 16 publications

References 32 publications

A novel strategy for D-psicose and lipase co-production using a co-culture system of engineered Bacillus subtilis and Escherichia coli and bioprocess analysis using metabolomics

A novel strategy for D-psicose and lipase co-production using a co-culture system of engineered Bacillus subtilis and Escherichia coli and bioprocess analysis using metabolomics

High-efficiency expression of the thermophilic lipase from Geobacillus thermocatenulatus in Escherichia coli and its application in the enzymatic hydrolysis of rapeseed oil

Biocatalysis at Extreme Temperatures: Enantioselective Synthesis of both Enantiomers of Mandelic Acid by Transesterification Catalyzed by a Thermophilic Lipase in Ionic Liquids at 120 °C

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