Characterization of cold adapted and ethanol tolerant β-glucosidase from Bacillus cellulosilyticus and its application for directed hydrolysis of cellobiose to ethanol

2018 ◽  
Vol 109 ◽  
pp. 872-879 ◽  
Author(s):  
Jian Wu ◽  
Alei Geng ◽  
Rongrong Xie ◽  
Hongcheng Wang ◽  
Jianzhong Sun
Keyword(s):  
Cold Adapted ◽  
Hydrolysis Of

scholarly journals Glucosylglycerate Biosynthesis in the Deepest Lineage of the Bacteria: Characterization of the Thermophilic Proteins GpgS and GpgP from Persephonella marina

2006 ◽  
Vol 189 (5) ◽  
pp. 1648-1654 ◽  
Author(s):  
Joana Costa ◽  
Nuno Empadinhas ◽  
Milton S. da Costa
Keyword(s):  
Escherichia Coli ◽  
High Temperatures ◽  
Gel Filtration ◽  
Maximal Activity ◽  
Thermophilic Bacterium ◽  
Cold Adapted ◽  
Dimeric Protein ◽  
Hydrolysis Of

ABSTRACT The pathway for the synthesis of glucosylglycerate (GG) in the thermophilic bacterium Persephonella marina is proposed based on the activities of recombinant glucosyl-3-phosphoglycerate (GPG) synthase (GpgS) and glucosyl-3-phosphoglycerate phosphatase (GpgP). The sequences of gpgS and gpgP from the cold-adapted bacterium Methanococcoides burtonii were used to identify the homologues in the genome of P. marina, which were separately cloned and overexpressed as His-tagged proteins in Escherichia coli. The recombinant GpgS protein of P. marina, unlike the homologue from M. burtonii, which was specific for GDP-glucose, catalyzed the synthesis of GPG from UDP-glucose, GDP-glucose, ADP-glucose, and TDP-glucose (in order of decreasing efficiency) and from d-3-phosphoglycerate, with maximal activity at 90°C. The recombinant GpgP protein, like the M. burtonii homologue, dephosphorylated GPG and mannosyl-3-phosphoglycerate (MPG) to GG and mannosylglycerate, respectively, yet at high temperatures the hydrolysis of GPG was more efficient than that of MPG. Gel filtration indicates that GpgS is a dimeric protein, while GpgP is monomeric. This is the first characterization of genes and enzymes for the synthesis of GG in a thermophile.

scholarly journals Cloning, Expression and Characterization of a Novel Cold-adapted β-galactosidase from the Deep-sea Bacterium Alteromonas sp. ML52

Marine Drugs ◽  
2018 ◽  
Vol 16 (12) ◽  
pp. 469 ◽  
Author(s):  
Jingjing Sun ◽  
Congyu Yao ◽  
Wei Wang ◽  
Zhiwei Zhuang ◽  
Junzhong Liu ◽  
...  
Keyword(s):  
Deep Sea ◽  
Glycoside Hydrolase ◽  
Sea Water ◽  
Maximum Activity ◽  
Glycoside Hydrolase Family ◽  
Cold Adapted ◽  
Hydrolysis Of ◽  
Hydrolase Family ◽  
Substrate Hydrolysis

The bacterium Alteromonas sp. ML52, isolated from deep-sea water, was found to synthesize an intracellular cold-adapted β-galactosidase. A novel β-galactosidase gene from strain ML52, encoding 1058 amino acids residues, was cloned and expressed in Escherichia coli. The enzyme belongs to glycoside hydrolase family 2 and is active as a homotetrameric protein. The recombinant enzyme had maximum activity at 35 °C and pH 8 with a low thermal stability over 30 °C. The enzyme also exhibited a Km of 0.14 mM, a Vmax of 464.7 U/mg and a kcat of 3688.1 S−1 at 35 °C with 2-nitrophenyl-β-d-galactopyranoside as a substrate. Hydrolysis of lactose assay, performed using milk, indicated that over 90% lactose in milk was hydrolyzed after incubation for 5 h at 25 °C or 24 h at 4 °C and 10 °C, respectively. These properties suggest that recombinant Alteromonas sp. ML52 β-galactosidase is a potential biocatalyst for the lactose-reduced dairy industry.

scholarly journals Pretreatment of Switchgrass for Production of Glucose via Sulfonic Acid-Impregnated Activated Carbon

Processes ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 504
Author(s):  
Yane Ansanay ◽  
Praveen Kolar ◽  
Ratna Sharma-Shivappa ◽  
Jay Cheng ◽  
Consuelo Arellano
Keyword(s):  
Activated Carbon ◽  
Sulfonic Acid ◽  
Solid Acid ◽  
Methanesulfonic Acid ◽  
Biofuel Production ◽  
Acid Catalysts ◽  
Effects Of Temperature ◽  
Carbon Catalysts ◽  
Hydrolysis Of

In the present research, activated carbon-supported sulfonic acid catalysts were synthesized and tested as pretreatment agents for the conversion of switchgrass into glucose. The catalysts were synthesized by reacting sulfuric acid, methanesulfonic acid, and p-toluenesulfonic acid with activated carbon. The characterization of catalysts suggested an increase in surface acidities, while surface area and pore volumes decreased because of sulfonation. Batch experiments were performed in 125 mL serum bottles to investigate the effects of temperature (30, 60, and 90 °C), reaction time (90 and 120 min) on the yields of glucose. Enzymatic hydrolysis of pretreated switchgrass using Ctec2 yielded up to 57.13% glucose. Durability tests indicated that sulfonic solid-impregnated carbon catalysts were able to maintain activity even after three cycles. From the results obtained, the solid acid catalysts appear to serve as effective pretreatment agents and can potentially reduce the use of conventional liquid acids and bases in biomass-into-biofuel production.

Characterization of endogenous endopeptidases and exopeptidases and application for the limited hydrolysis of peanut proteins

2021 ◽  
Vol 345 ◽  
pp. 128764
Author(s):  
Yeming Chen ◽  
Hongsheng Zhang ◽  
Caimeng Zhang ◽  
Xiangzhen Kong ◽  
Yufei Hua
Keyword(s):  
Peanut Proteins ◽  
Hydrolysis Of

scholarly journals Multifunctional Biodegradable Vascular PLLA Scaffold with Improved X-ray Opacity, Anti-Inflammation, and Re-Endothelization

Polymers ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 1979
Author(s):  
Ho In Lee ◽  
Yun Heo ◽  
Seung-Woon Baek ◽  
Da-Seul Kim ◽  
Duck Hyun Song ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Inflammatory Responses ◽  
Neointimal Hyperplasia ◽  
Film Surface ◽  
High Mechanical Property ◽  
X Ray ◽  
Stent Restenosis ◽  
X Ray Computed ◽  
Hydrolysis Of

Poly(L-lactic acid) (PLLA) has been used as a biodegradable vascular scaffold (BVS) material due to high mechanical property, biodegradability, and biocompatibility. However, acidic byproducts from hydrolysis of PLLA reduce the pH after the surrounding implanted area and cause inflammatory responses. As a result, severe inflammation, thrombosis, and in-stent restenosis can occur after implantation by using BVS. Additionally, polymers such as PLLA could not find on X-ray computed tomography (CT) because of low radiopacity. To this end, here, we fabricated PLLA films as the surface of BVS and divided PLLA films into two coating layers. At the first layer, PLLA film was coated by 2,3,5-triiodobenzoic acid (TIBA) and magnesium hydroxide (MH) with poly(D,L-lactic acid) (PDLLA) for radiopaque and neutralization of acidic environment, respectively. The second layer of coated PLLA films is composed of polydopamine (PDA) and then cystamine (Cys) for the generation of nitric oxide (NO) release, which is needed for suppression of smooth muscle cells (SMCs) and proliferation of endothelial cells (ECs). The characterization of the film surface was conducted via various analyses. Through the surface modification of PLLA films, they have multifunctional abilities to overcome problems of BVS effectively such as X-ray penetrability, inflammation, thrombosis, and neointimal hyperplasia. These results suggest that the modification of biodegradable PLLA using TIBA, MH, PDA, and Cys will have important potential in implant applications.

scholarly journals Crystal structure of thermostable alkylsulfatase SdsAP from Pseudomonas sp. S9

2017 ◽  
Vol 37 (3) ◽  
Author(s):  
Lifang Sun ◽  
Pu Chen ◽  
Yintao Su ◽  
Zhixiong Cai ◽  
Lingwei Ruan ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Sodium Dodecyl ◽  
Titration Calorimetry ◽  
Pseudomonas Sp ◽  
Sterol Carrier Protein ◽  
Dimerization Domain ◽  
Alkyl Chains ◽  
Hydrolysis Of

A novel alkylsulfatase from bacterium Pseudomonas sp. S9 (SdsAP) was identified as a thermostable alkylsulfatases (type III), which could hydrolyze the primary alkyl sulfate such as sodium dodecyl sulfate (SDS). Thus, it has a potential application of SDS biodegradation. The crystal structure of SdsAP has been solved to a resolution of 1.76 Å and reveals that SdsAP contains the characteristic metallo-β-lactamase-like fold domain, dimerization domain, and C-terminal sterol carrier protein type 2 (SCP-2)-like fold domain. Kinetic characterization of SdsAP to SDS by isothermal titration calorimetry (ITC) and enzymatic activity assays of constructed mutants demonstrate that Y246 and G263 are important residues for its preference for the hydrolysis of ‘primary alkyl’ chains, confirming that SdsAP is a primary alkylsulfatase.

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