• Title/Summary/Keyword: Gel chromatography

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Production of Poly (3-Hydroxybutyrate-co-3-Hydroxyvalerate) by Bacillus sp. EMK-5020 Using Makgeolli Lees Enzymatic Hydrolysate and Propionic Acid as Carbon Sources (막걸리 주박 가수분해 산물과 propionic acid를 탄소원으로 이용한 Bacillus sp. EML-5020 균주로부터 poly (3-hydroxybutyrate-co-3-hydroxyvalerate) 생합성)

  • Kwon, Kyungjin;Kim, Jong-Sik;Chung, Chung-Wook
    • Journal of Life Science
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    • v.32 no.7
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    • pp.510-522
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    • 2022
  • In this study, to biosynthesize PHA with properties more similar to polypropylene, a Bacillus sp. EMK-5020 strain that biosynthesized poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was isolated from soil. Bacillus sp. EMK-5020 strain biosynthesized PHBV containing 1.3% 3-hydroxyvalerate (3HV) using reducing sugar contained in Makgeolli lees enzymatic hydrolysate (MLEH) as a single carbon source. As the amount of propionic acid, which was added as a second carbon source, increased, the content of 3HV also increased. PHBV containing up to 48.6% of 3HV was synthesized when 1.0 g/l of propionic acid was added. Based on these results, the strain was cultured for 72 hr in a 3 l fermenter using reducing sugar in MLEH (20 g/l) and propionic acid (1 g/l) as the main and secondary carbon sources, respectively. As a result, 6.4 g/l DCW and 50 wt% of PHBV (MLEH-PHBV) containing 8.9% 3HV were biosynthesized. Through gel permeation chromatography and thermogravimetric analysis, it was confirmed that the average molecular weight and the decomposition temperature of MLEH-PHBV were 152 kDa and 273℃, respectively. In conclusion, the Bacillus sp. EMK-5020 strain could biosynthesize PHBV containing various 3HV fractions when MLEH and propionic acid were used as carbon sources, and PHBV-MLEH containing 8.9% 3HV was confirmed to have higher thermal stability than standard PHBV (8% 3HV).

Glycoprotein in the Fruit Body of Sarcodon aspratus (능이자실체의 Glycoprotein)

  • Cho, Nam-Seok;Choi, Tae-Ho;Cho, Hee-Yeon;Leonowicz, Andrzej
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.5
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    • pp.51-58
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    • 2004
  • This study was performed to investigate compositions of inorganic elements, amino acids and glycoprotein fractions as biological substances in fruit body of Sarcodon aspratus. The fruit body of Sarcodon aspratus contained Ca, Mg, Zn, Mn, Fe, Cu, and Pb, in particular high Ca and Na. Hot water extracts consisted of 54% of polysaccharide fraction and 32.6% of protein. In amino acids composition, fourteen free amino acids were detected, mainly glutamic acid, alanine and arginine. Fifteen kinds of total amino acids were contained with major components of glutamic acid, aspartic acid, serine and threonine. Concerned to glycoprotein extraction, 95% ethyl alcohol concentration gave the highest yields with 70.6% sugar fraction, 332% glycoprotein. Different ethyl alcohol concentration resulted in different protein precipitations, and lower concentration ethyl alcohol in the range of 30 to 70% gave more than 92% of higher sugar fraction. Crude glycoprotein (GP) was fractionated by P fraction of more than MW 300,000, P-1 fraction unadsorbed by DEAE-Sephadex, P-2 fractionated from P-1 by Sepharose 2B gel chromatography and P-3 fraction adsorbed by DEAE-Sephadex. Total sugars were increased and protein contents decreased during fractionation. GP and P-3 contained glucose, galactose, mannose and fucose. GP had high glucose with high contents of glutamic acid, serine, alanine and glycine. P-3 fraction contained high mannose with aspartic acid, glutamic acid, and glycine. P-2 fraction was 700,000 MW with high glucose and fucose, and low protein of 1.1%, high amounts of aspartic acid, glutamic acid and alanine, but no mannose and no cysteine.

Cell Migration and Wound Healing Activities of Recombinant Thymosin β-4 Expressed in Escherichia coli (재조합 Thymosin β-4의 세포이동능과 상처치유능)

  • Hong, Kyo-Chang;Choi, Yung Hyun;Kim, Gun-Do;Cha, Hee-Jae;Jeon, Sung-Jong;Nam, Soo-Wan
    • Journal of Life Science
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    • v.32 no.2
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    • pp.135-141
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    • 2022
  • Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein - chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.

The synthesis of dextran from rice hydrolysates using Gluconobacter oxydans KACC 19357 bioconversion (Gluconobacter oxydans 생물전환을 통한 쌀 가수분해물 유래 dextran 합성)

  • Seung-Min Baek;Hyun Ji Lee;Legesse Shiferaw Chewaka;Chan Soon Park;Bo-Ram Park
    • Food Science and Preservation
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    • v.31 no.1
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    • pp.149-160
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    • 2024
  • Dextran is a glucose homo-polysaccharide with a predominantly α-1,6 glycosidic linkage of microbial source and is known to be produced primarily by lactic acid bacteria. However, it can also be obtained through the dextran dextrinase of acetic acid bacteria (Gluconobacter oxydans). The dextrin-based dextran was obtained from rice starch using G. oxydans fermentation of rice hydrolysate, and its properties were studied. Both dextrin- and rice hydrolysate-added media maintained the OD value of 6 after 20 h of incubation with acetic acid bacteria, and the gel permeation chromatography (GPC) analysis of the supernatant after 72 h of incubation confirmed that a polymeric material with DP of 480 and 405, which was different from the composition of the substrate in the medium, was produced. The glucose linkage pattern of the polysaccharide was confirmed using the proton nuclear magnetic resonance (1H-NMR) and the increased α-1,4:α-1,6 bond ratio from 0.23 and 0.13 to 1:2.37 and 1:4.4, respectively, indicating that the main bonds were converted to α-1,6 bonds. The treatment of dextrin with a rat-derived alpha-glucosidase digestive enzyme resulted in a slow release of glucose, suggesting that rice hydrolysate can be converted to dextran using acetic acid bacteria with glycosyltransferase activity to produce high-value bio-materials with slowly digestible properties.