Molecular Characterization of a Novel Family VIII Esterase with β-Lactamase Activity (PsEstA) from Paenibacillus sp.

The immobilization of the biocatalysts onto magnetic nanoparticles (MNPs) has been extensively applied since the external magnetic field facilitates the enzyme recovery from the reaction mixture. In the present study, glutaraldehyde-modified magnetite-cornstarch nanoparticles (MCNs) were successfully synthesized, elaborately characterized by ZetaSizer and surface-enhanced raman spectroscopy (SERS), and used for the immobilization of a thermoalkalophilic esterase from Geobacillus sp. The optimal immobilization conditions were obtained at 65oC, 2:3 molar ratios of Fe2+:Fe3+ and 1 g cornstarch resulted in approximately 90 nm magnetic particles in size. Also, immobilization yield and entrapment efficiency of the esterase were found as 74% and 82%, respectively. Scanning Electron Microscopy (SEM) micrographs showed that MCNs were uniform, spherical in shape, and well dispersed and esterase immobilized MCNs displayed similar morphology as free MCNs. The maximum activity of free and immobilized esterase was obtained at 80oC and pH 9. Immobilization onto glutaraldehyde-modified MCNs significantly enhanced the esterase thermostability. Additionally, the immobilized esterase kept its residual activity of 75% after three sequential cycles, suggesting that it has favorable operational stability.

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“…7). In literature, there have been several reports acquiring similar ndings to the present study [36,[47][48][49].…”

Section: Resultssupporting

confidence: 87%

“…Similar ndings have been reported about immobilized esterases on various support materials including MNPs. They have possessed a residual activity of above 70% next three sequential cycles [35,[47][48][49][50][51].…”

Section: Resultsmentioning

confidence: 99%

Immobilization of Thermoalkalophilic Esterase Onto Magnetic-Cornstarch Nanoparticle

Öz

Sürmeli

Şanlı‐Mohamed

2021

Preprint

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“…(CLEAs-PsEstA). 55 The activity of another immobilized esterase from Neisseria meningitides on crosslinked enzyme aggregates (NmSGNH1-CLEAs) was slightly reduced by Triton X-100; however, it was dramatically dropped by SDS. 53 In another study, SDS and Triton X-100 dramatically reduced immobilized esterase Lx-Est BAS ΔSP activity by 80% and 62%, respectively.…”

Section: Effect Of Chemicalsmentioning

confidence: 99%

“…9 There have been some recent reports with similar results on immobilized esterases using different techniques, showing at least 70% of residual activity after three sequential cycles. 45,47,53,55,59,60 The influence of bead size The effect of bead sizes (1.8, 2.6, and 3.9 mm) was investigated on the activity of the immobilized enzyme. This analysis results showed that the highest esterase activity was determined at 1.8 mm diameter beads the smallest size obtained in the present study (Fig.…”

Section: Kinetic Studiesmentioning

confidence: 99%

Thermoalkalophilic recombinant esterase entrapment in chitosan/calcium/alginate‐blended beads and its characterization

Tercan

Sürmeli

Şanlı‐Mohamed

2021

J of Chemical Tech & Biotech

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BACKGROUND: Esterases (EC 3.1.1.1), a class of hydrolases, degrade the ester bonds of lipids into alcohol and carboxylic acids and synthesize carboxylic ester bonds. They are used in a variety of biotechnological, industrial, environmental, and pharmaceutical applications due to their many valuable properties. Particularly, extremophilic esterases with many superior properties are of great interest for various reactions. Immobilization of enzymes may provide some advantages over free enzymes not only to improve the properties of enzymes but also to increase the reusability of biocatalyst in industrial applications. Therefore, many different immobilization applications for enzymes have been reported in various studies. To our knowledge, a thermophilic esterase has not so far been immobilized by entrapment using chitosan/calcium/alginate-blended beads. Here, we reported the immobilization of thermoalkalophilic recombinant esterase by entrapment using chitosan/calcium/alginateblended beads, and then the entrapped esterase was characterized biochemically in details.RESULTS: In the present study, a thermophilic recombinant esterase was immobilized by entrapment in chitosan/calcium/alginateblended beads for the first time. The 0.5 mg mL −1 purified recombinant esterase was entrapped in 1% chitosan, 2% alginate, and 0.7 M CaCl 2 blended beads. The results showed that immobilization yield and entrapment efficiency of the entrapped esterase were 69.5% and 80.4%, respectively. SEM micrograph showed that the surface of the beads resembled a mesh and very compact structures. Chitosan/calcium/alginate-blended beads exhibited an 18.8% swelling ratio and had a moderate porous structure. The entrapment technique highly enhanced the thermostability of the esterase and shifted its optimum temperature from 65 to 80°C. The immobilized esterase was very stable in a wide range of pH (8.5-11) displaying maximum activity at pH 9. ZnCl 2 slightly increased the activity of immobilized esterase whereas several metal ions reduced the enzyme activity. When the enzyme was immobilized in chitosan/calcium/alginate-blended beads, its K m increased about 2 times and V max value decreased almost 1.5 times. Immobilization allowed repeated uses of the esterase having good operational stability in a continuous process. CONCLUSION:The results revealed that the immobilization of a thermophilic recombinant esterase by entrapment in chitosan/ calcium/alginate-blended beads exhibited considerably better compared to other immobilization processes with various entrapment strategies.

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“…Substrate specificities of HaSGNH1 and its variants were determined using p-nitrophenyl (p-NP) esters and naphthyl ester derivatives [54]. The standard assay solution included 50 μM substrates in 20 mM Tris-HCl (pH 8.0) with 0.5 μg HaSGNH1, and the assay was run for 30 s at 25 °C.…”

Section: Biochemical Characterization Of Hasgnh1mentioning

confidence: 99%

Biodiesel and flavor compound production using a novel promiscuous cold-adapted SGNH-type lipase (HaSGNH1) from the psychrophilic bacterium Halocynthiibacter arcticus

Yoo

Jeon

et al. 2020

Biotechnol Biofuels

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Background: Biodiesel and flavor compound production using enzymatic transesterification by microbial lipases provides mild reaction conditions and low energy cost compared to the chemical process. SGNH-type lipases are very effective catalysts for enzymatic transesterification due to their high reaction rate, great stability, relatively small size for convenient genetic manipulations, and ease of immobilization. Hence, it is highly important to identify novel SGNH-type lipases with high catalytic efficiencies and good stabilities. Results: A promiscuous cold-adapted SGNH-type lipase (HaSGNH1) from Halocynthiibacter arcticus was catalytically characterized and functionally explored. HaSGNH1 displayed broad substrate specificity that included tert-butyl acetate, glucose pentaacetate, and p-nitrophenyl esters with excellent stability and high efficiency. Important amino acids (N83, M86, R87, F131, and I173F) around the substrate-binding pocket were shown to be responsible for catalytic activity, substrate specificity, and reaction kinetics. Moreover, immobilized HaSGNH1 was used to produce high yields of butyl and oleic esters. Conclusions: This work provides a molecular understanding of substrate specificities, catalytic regulation, immobilization, and industrial applications of a promiscuous cold-adapted SGNH-type lipase (HaSGNH1) from H. arcticus. This is the first analysis on biodiesel and flavor synthesis using a cold-adapted halophilic SGNH-type lipase from a Halocynthiibacter species.

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Molecular Characterization of a Novel Family VIII Esterase with β-Lactamase Activity (PsEstA) from Paenibacillus sp.

Cited by 11 publications

References 51 publications

Immobilization of Thermoalkalophilic Esterase Onto Magnetic-Cornstarch Nanoparticle

Immobilization of Thermoalkalophilic Esterase Onto Magnetic-Cornstarch Nanoparticle

Thermoalkalophilic recombinant esterase entrapment in chitosan/calcium/alginate‐blended beads and its characterization

Biodiesel and flavor compound production using a novel promiscuous cold-adapted SGNH-type lipase (HaSGNH1) from the psychrophilic bacterium Halocynthiibacter arcticus

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