• Title/Summary/Keyword: microbial cellulose

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Screening and Characterization of an Enzyme with ${\beta}-Glucosidase$ Activity from Environmental DNA

  • Kim, Soo-Jin;Lee, Chang-Muk;Kim, Min-Young;Yeo, Yun-Soo;Yoon, Sang-Hong;Kang, Han-Cheol;Koo, Bon-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.905-912
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    • 2007
  • A novel ${\beta}-glucosidase$ gene, bglA, was isolated from uncultured soil bacteria and characterized. Using genomic libraries constructed from soil DNA, a gene encoding a protein that hydrolyzes a fluorogenic analog of cellulose, 4-methylumbelliferyl ${\beta}-D-cellobioside$ (MUC), was isolated using a microtiter plate assay. The gene, bglA, was sequenced using a shotgun approach, and expressed in E. coli. The deduced 55-kDa amino acid sequence for bglA showed a 56% identity with the family 1 glycosyl hydrolase Chloroflexus aurantiacus. BglA included two conserved family 1 glycosyl hydrolase regions. When using $p-nitrophenyl-{\beta}-D-glucoside$ (pNPG) as the substrate, the maximum activity of the purified ${\beta}-glucosidase$ exhibited at pH 6.5 and $55^{\circ}C$, and was enhanced in the presence of $Mn^{2+}$. The $K_m\;and\;V_{max}$ values for the purified enzyme with pNPG were 0.16 mM and $19.10{\mu}mol/min$, respectively. The purified BglA enzyme hydrolyzed both pNPG and $p-nitrophenyl-{\beta}-D-fucoside$. The enzyme also exhibited substantial glycosyl hydrolase activities with natural glycosyl substrates, such as sophorose, cellobiose, cellotriose, cellotetraose, and cellopentaose, yet low hydrolytic activities with gentiobiose, salicin, and arbutin. Moreover, BglA was able to convert the major ginsenoside $Rb_1$ into the pharmaceutically active minor ginsenoside Rd within 24 h.

Characterization of a Thermophilic Lignocellulose-Degrading Microbial Consortium with High Extracellular Xylanase Activity

  • Zhang, Dongdong;Wang, Yi;Zhang, Chunfang;Zheng, Dan;Guo, Peng;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.305-313
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    • 2018
  • A microbial consortium, TMC7, was enriched for the degradation of natural lignocellulosic materials under high temperature. TMC7 degraded 79.7% of rice straw during 15 days of incubation at $65^{\circ}C$. Extracellular xylanase was effectively secreted and hemicellulose was mainly degraded in the early stage (first 3 days), whereas primary decomposition of cellulose was observed as of day 3. The optimal temperature and initial pH for extracellular xylanase activity and lignocellulose degradation were $65^{\circ}C$ and between 7.0 and 9.0, respectively. Extracellular xylanase activity was maintained above 80% and 85% over a wide range of temperature ($50-75^{\circ}C$) and pH values (6.0-11.0), respectively. Clostridium likely had the largest contribution to lignocellulose conversion in TMC7 initially, and Geobacillus, Aeribacillus, and Thermoanaerobacterium might have also been involved in the later phase. These results demonstrate the potential practical application of TMC7 for lignocellulosic biomass utilization in the biotechnological industry under hot and alkaline conditions.

Metagenomic and Proteomic Analyses of a Mangrove Microbial Community Following Green Macroalgae Enteromorpha prolifera Degradation

  • Wu, Yijing;Zhao, Chao;Xiao, Zheng;Lin, Hetong;Ruan, Lingwei;Liu, Bin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2127-2137
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    • 2016
  • A mangrove microbial community was analyzed at the gene and protein levels using metagenomic and proteomic methods with the green macroalgae Enteromorpha prolifera as the substrate. Total DNA was sequenced on the Illumina HiSeq 2000 PE-100 platform. Two-dimensional gel electrophoresis in combination with liquid chromatography tandem mass spectrometry was used for proteomic analysis. The metagenomic data revealed that the orders Pseudomonadales, Rhizobiales, and Sphingomonadales were the most prevalent in the mangrove microbial community. By monitoring changes at the functional level, proteomic analyses detected ATP synthase and transporter proteins, which were expressed mainly by members of the phyla Proteobacteria and Bacteroidetes. Members of the phylum Proteobacteria expressed a high number of sugar transporters and demonstrated specialized and efficient digestion of various glycans. A few glycoside hydrolases were detected in members of the phylum Firmicutes, which appeared to be the main cellulose-degrading bacteria. This is the first report of multiple "omics" analysis of E. prolifera degradation. These results support the fact that key enzymes of glycoside hydrolase family were expressed in large quantities, indicating the high metabolic activity of the community.

Studies on the Development of a Microbial Cryoprotectant Formulation Using a W/O/W Multiple Emulsion System

  • Bae, Eun-Kyung;Cho, Young-Hee;Park, Ji-Yong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.673-679
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    • 2004
  • A microbial cryoprotectant formulation using a W/O/W multiple emulsion system was developed. The psychrotolerant microorganism, B4, isolated from soil in South Korea, was observed by the drop freezing method, in which the microorganism sample inhibited ice nucleation activity. The antifreeze activity was eliminated when the microorganism sample was treated with protease, indicating that the antifreeze activity was due to the presence of antifreeze protein. The result of the l6S rDNA sequencing indicated the B4 strain was most closely related to a species of the genus Bacillus. Culture broth of B4 strain (Bacillus sp.) and rapeseed oil containing 1 % polyglycerine polyricinolate (PGPR) were used as core and wall material, respectively. The most stable W/O emulsion was prepared at a core/oil ratio of 1:2. The highest W/O/W emulsion stability was achieved when the primary emulsion to external aqueous phase containing 0.5% caster oil polyoxyethylene ether $(COG25^{TM})$ ratio was 1:1. Microcrystalline cellulose showed better W/O/W emulsion stability than other polymer types. The viability of cells in a W/O/W emulsion was higher than free cells during storage at $37^\circ{C}$. An acidic pH and UV exposure decreased the viability of free cells, but cells in W/O/W emulsion were more stable under these conditions.

Isolation and characterization of microorganisms biological damage of Dongchundang (동춘당 생물학적 가해 미생물의 분리 및 특성)

  • Lee, Jeung-Min;Kim, Young-Hee;Hong, Jin-Young;Jo, Chang-Wook;Kim, Soo Ji;Seo, Min Seok
    • 보존과학연구
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    • s.35
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    • pp.111-119
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    • 2014
  • Microorganisms were isolated from Dongchundang(wooden cultural heritage) with PDA medium culture. Nineteen species shows the cellulolytic activity. Methylobacterium sp. was the most active in cellulose degradation. The growth curve and pH were measured during incubation of the microorganism for 72 hours. The pH was increased with the increasing of microbial growth. The degree of cellulose degradation was determined with the amount of reducing sugar by use of dinitrosalicylic acid (DNS) method. The amount of reducing sugar was decreased after 45 hours. As a results, It should suggested that wood component were deteriorated by Methylobacterium sp..

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Isolation of cellulosic biomass degrading microorganisms from different sources for low cost biofuel production

  • Sheikh, M. Mominul Islam;Kim, Chul-Hwan;Lee, Ji-Yong;Yeasmin, Shabina;Park, Hyeon-Jin;Kim, Gyeong-Chul;Kim, Sung-Ho;Kim, Jae-Won
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2011.04a
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    • pp.81-91
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    • 2011
  • Current fuel ethanol research and development deals with process engineering trends for improving biotechnological production of ethanol. Recently, a large amount of studies regarding the utilization of lignocellulosic biomass as a good feedstock for producing fuel ethanol is being carried out worldwide. The plant biomass is mainly composed of cellulose, hemicellulose and lignin. The main challenge in the conversion of biomass into ethanol is the complex, rigid and harsh structures which require efficient process and cost effective to break down. The isolation of microorganisms is one of the means for obtaining enzymes with properties suitable for industrial applications. For these reasons, crude cultures containing cellulosic biomass degrading microorganisms were isolated from rice field soil, cow farm soil and rotten rice straw from cow farm. Carboxymethyl cellulose (CMC), xylan and Avicel (microcrystalline cellulose) degradation zone of clearance on agar platefrom rice field soil resulted approximately at 25 mm, 24 mm and 22 mm respectively. As for cow farm soil, CMC, xylan and Avicel degradation clearancezone on agar plate resulted around at 24mm, 23mm and 21 mm respectively. Rotten rice straw from cow farm also resulted for CMC, xylan and Avicel degradation zone almost at 24 mm, 23 mm and 22 mm respectively. The objective of this study is to isolatebiomass degrading microbial strains having good efficiency in cellulose hydrolysis and observed the effects of different substrates (CMC, xylan and Avicel) on the production of cellulase enzymes (endo-glucanase, exo-glucanase, cellobiase, xylanase and avicelase) for producing low cost biofuel from cellulosic materials.

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Production of Bacterial Cellulose by Gluconacetobacter hansenii Using a New Bioreactor Equipped with Centrifugal Impellers (원심 임펠러가 장착된 발효조에서 G. hansenii에 의한 미생물셀룰로오스 생산)

  • Khan, Salman;Shehzad, Omer;Khan, Taous;Ha, Jung Hwan;Park, Joong Kon
    • Korean Chemical Engineering Research
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    • v.47 no.4
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    • pp.506-511
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    • 2009
  • In order to improve the bacterial cellulose(BC) production yield, centrifugal and inclined centrifugal impellers were developed. A 6 flat-blade turbine impeller was used as a control system. The flow pattern in the fermenter and volumetric oxygen transfer coefficient($k_La$) of these fermentation systems were studied. Fermentations were carried out for the production of BC by G. hansenii PJK in a 2-L jar fermenter equipped with new impellers. Liquid medium was circulated from the bottom, through the cylinder of the impeller and to the wall. The volumetric oxygen transfer coefficients, $k_La$, of inclined centrifugal and centrifugal impeller systems at 100 rpm were 23 and 15% of the conventional turbine impeller system, respectively. However, the conversion of microbial cells to cellulose non-producing mutant decreased and this results in the increase in BC production at low rotating speed of impellers.

Shelf-life prediction of fresh ginseng packaged with plastic films based on a kinetic model and multivariate accelerated shelf-life testing

  • Jong-Jin Park;Jeong-Hee Choi;Kee-Jai Park;Jeong-Seok Cho;Dae-Yong Yun;Jeong-Ho Lim
    • Food Science and Preservation
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    • v.30 no.4
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    • pp.573-588
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    • 2023
  • The purpose of this study was to monitor changes in the quality of ginseng and predict its shelf-life. As the storage period of ginseng increased, some quality indicators, such as water-soluble pectin (WSP), CDTA-soluble pectin (CSP), cellulose, weight loss, and microbial growth increased, while others (Na2CO3-soluble pectin/NSP, hemicellulose, starch, and firmness) decreased. Principal component analysis (PCA) was performed using the quality attribute data and the principal component 1 (PC1) scores extracted from the PCA results were applied to the multivariate analysis. The reaction rate at different temperatures and the temperature dependence of the reaction rate were determined using kinetic and Arrhenius models, respectively. Among the kinetic models, zeroth-order models with cellulose and a PC1 score provided an adequate fit for reaction rate estimation. Hence, the prediction model was constructed by applying the cellulose and PC1 scores to the zeroth-order kinetic and Arrhenius models. The prediction model with PC1 score showed higher R2 values (0.877-0.919) than those of cellulose (0.797-0.863), indicating that multivariate analysis using PC1 score is more accurate for the shelf-life prediction of ginseng. The predicted shelf-life using the multivariate accelerated shelf-life test at 5, 20, and 35℃ was 40, 16, and 7 days, respectively.

Enhancing the Anaerobic Digestion of Corn Stalks Using Composite Microbial Pretreatment

  • Yuan, Xufeng;Li, Peipei;Wang, Hui;Wang, Xiaofen;Cheng, Xu;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.746-752
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    • 2011
  • A composite microbial system (XDC-2) was used to pretreat and hydrolyze corn stalk to enhance anaerobic digestion. The results of pretreatment indicated that sCOD concentrations of hydrolysate were highest (8,233 mg/l) at the fifth day. XDC-2 efficiently degraded the corn stalk by nearly 45%, decreasing the cellulose content by 22.7% and the hemicellulose content by 74.1%. Total levels of volatile products peaked on the fifth day. The six major compounds present were ethanol (0.29 g/l), acetic acid (0.55 g/l), 1,2-ethanediol (0.49 g/l), propionic acid (0.15 g/l), butyric acid (0.22 g/l), and glycerine (2.48 g/l). The results of anaerobic digestion showed that corn stalks treated by XDC-2 produced 68.3% more total biogas and 87.9% more total methane than untreated controls. The technical digestion time for the treated corn stalks was 35.7% shorter than without treatment. The composite microbial system pretreatment could be a cost-effective and environmentally friendly microbial method for efficient biological conversion of corn stalk into bioenergy.

Property Changes of the Salt-Seasoned and Fermented the Broken Roes of Alaska Pollock Stuffed into Cellulose Casing (Cellulose casing에 충진한 명태 절란젓의 숙성중 품질변화)

  • Park, Jong-Hyuk;Kim, Sang-Moo
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.220-224
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    • 2002
  • Alaska pollock roe is mainly used for the production of salt-seasoned and fermented seafood (Myungran-jeot). Alaska pollock roes with broken egg membrane are usually discarded as a waste product. In this study, the broken roes of Alaska pollock were salt-seasoned and stuffed into cellulose casing for commercial production. The chemical and microbial changes of the broken roes of Alaska pollock stuffed into cellulose casing fermented at 5 and $25^{\circ}C$, respectively, were analyzed at different ripening periods. On 5 week fermentation, pH decreased down to 5.60 and 5.10 at 5 and $25^{\circ}C$, respectively, but the amounts of lactic acid, amino-nitrogen, and volatile basic nitrogen increased continously as ripening period increased, higher at 25 than $5^{\circ}C$. The amounts of amino-nitrogen, 620 and 780 mg/100 g, were the highest on 3 week fermentation at $5^{\circ}C$ and on 1 week at $25^{\circ}C$, respectively. The numbers of total viable cell and lactic acid bacteria, $3.1{\times}10^6$ and $3.1{\times}10^5\;CFU/g$ at $5^{\circ}C$, and $1.9{\times}10^7$ and $2.8{\times}10^6\;CFU/g$ at $25^{\circ}C$, respectively, were the highest on 2 week fermentation.