• Genomics & Informatics
• /
• v.11 no.3
• /
• pp.114-120
• /
• 2013

The microbial diversity in soil ecosystems is higher than in any other microbial ecosystem. The majority of soil microorganisms has not been characterized, because the dominant members have not been readily culturable on standard cultivation media; therefore, the soil ecosystem is a great reservoir for the discovery of novel microbial enzymes and bioactivities. The soil metagenome, the collective microbial genome, could be cloned and sequenced directly from soils to search for novel microbial resources. This review summarizes the microbial diversity in soils and the efforts to search for microbial resources from the soil metagenome, with more emphasis on the potential of bioprospecting metagenomics and recent discoveries.

• Korean Journal of Environmental Agriculture
• /
• v.19 no.5
• /
• pp.375-381
• /
• 2000

Quantifying the changes of soil microbial biomass and activity of enzymes are important to understand the dynamics of active soil C and N pools. The dynamics of soil microbial biomass C and N and the activity of enzymes over entire growth period of soybean-(Glycine max (L) Merr.)-wheat (Triticum aestivum L.) sequence on a Typic Haplustert as influenced by organic manure and inorganic fertilizer N were investigated in a field experiment. The application of farmyard manure at 4 to 16 $Mg{\cdot}ha^{-1}\;y^{-1}r^{-1}$ along with fertilizer nitrogen at 50 or 180 $kg{\cdot}ha^{-1}$ increased the mean soil microbial biomass from 1.12 to 2.05 fold over unmanured soils under soybean-wheat system. Irrespective of organic and chemical fertilizer N application, the soil microbial biomass was maximum during the first two months at active growing stage of the crops and subsequently declined with crop maturity. The mean annual microbial activity was significantly increased when manure and chemical fertilizer at 8 $Mg{\cdot}ha^{-1}$ and 50/180 N $kg{\cdot}ha^{-1}$, respectively were applied. The C turnover rate decreased by 47 to 72 % when the level of farmyard manure was increased from 4 to 8 and 16 $Mg{\cdot}ha^{-1}$. There were significant correlations between biomass C, available N, dehydrogenase, phosphatase and yield of the crops.

• Microbiology and Biotechnology Letters
• /
• v.48 no.4
• /
• pp.515-524
• /
• 2020

With the increasing demand for enzymes in industrial applications there is a growing need to easily produce industrially important microbial enzymes. This study was carried out to screen the indigenous refinery bacterial isolates for their production of two industrially important enzymes i.e. lipase and protease. A total of 15 bacterial strains were isolated using Soil Extract Agar media from the oil-contaminated environment and one was shown to produce high quality lipase and protease enzymes. The culture conditions (culture duration, temperature, source of nitrogen, carbon, and pH) were optimized to produce the optimum amount of both the lipase (37.6 ± 0.2 Uml-1) and the protease (41 ± 0.4 Uml-1) from this isolate. Productivity of both enzymes was shown to be maximized at pH 7.5 in a medium containing yeast extract and peptone as nitrogen sources and sucrose and galactose as carbon sources when incubated at 35 ± 1℃ for 48 h. Bacterial strain SAB06 was identified as Bacillus licheniformis (MT250345) based on biochemical, morphological, and molecular characteristics. Further studies are required to evaluate and optimize the purification and characterization of these enzymes before they can be recommended for industrial or environmental applications.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.5
• /
• pp.740-746
• /
• 2021

Efficient cellulolytic enzyme production is important for the development of lignocellulose-degrading enzyme mixtures. However, purification of cellulases from their native hosts is time- and labor-consuming. In this study, a constitutive expression system was developed in Penicillium oxalicum for the secreted production of proteins. Using a constitutive polyubiquitin gene promoter and cultivating with glucose as the sole carbon source, nine cellulolytic enzymes of different origins with relatively high purity were produced within 48 h. When supplemented to a commercial cellulase preparation, cellobiohydrolase I from P. funiculosum and cellobiohydrolase II from Talaromyces verruculosus showed remarkable enhancing effects on the hydrolysis of steam-exploded corn stover. Additionally, a synergistic effect was observed for these two cellobiohydrolases during the hydrolysis. Taken together, the constitutive expression system provides a convenient tool for the production of cellulolytic enzymes, which is expected to be useful in the development of highly efficient lignocellulose-degrading enzyme mixtures.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.8
• /
• pp.1123-1133
• /
• 2021

Biodegradation is the key process involved in natural lignocellulose biotransformation and utilization. Microbial consortia represent promising candidates for applications in lignocellulose conversion strategies for biofuel production; however, cooperation among the enzymes and the labor division of microbes in the microbial consortia remains unclear. In this study, metagenomic analysis was performed to reveal the community structure and extremozyme systems of a lignocellulolytic microbial consortium, TMC7. The taxonomic affiliation of TMC7 metagenome included members of the genera Ruminiclostridium (42.85%), Thermoanaerobacterium (18.41%), Geobacillus (10.44%), unclassified_f__Bacillaceae (7.48%), Aeribacillus (2.65%), Symbiobacterium (2.47%), Desulfotomaculum (2.33%), Caldibacillus (1.56%), Clostridium (1.26%), and others (10.55%). The carbohydrate-active enzyme annotation revealed that TMC7 encoded a broad array of enzymes responsible for cellulose and hemicellulose degradation. Ten glycoside hydrolases (GHs) endoglucanase, 4 GHs exoglucanase, and 6 GHs β-glucosidase were identified for cellulose degradation; 6 GHs endo-β-1,4-xylanase, 9 GHs β-xylosidase, and 3 GHs β-mannanase were identified for degradation of the hemicellulose main chain; 6 GHs arabinofuranosidase, 2 GHs α-mannosidase, 11 GHs galactosidase, 3 GHs α-rhamnosidase, and 4 GHs α-fucosidase were identified as xylan debranching enzymes. Furthermore, by introducing a factor named as the contribution coefficient, we found that Ruminiclostridium and Thermoanaerobacterium may be the dominant contributors, whereas Symbiobacterium and Desulfotomaculum may serve as "sugar cheaters" in lignocellulose degradation by TMC7. Our findings provide mechanistic profiles of an array of enzymes that degrade complex lignocellulosic biomass in the microbial consortium TMC7 and provide a promising approach for studying the potential contribution of microbes in microbial consortia.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2004.06a
• /
• pp.215-218
• /
• 2004

Dextrans make up a class of polysaccharides that are D-glucans of various structures with contiguous $\alpha$-1longrightarrow6 ~6 glycosidic linkages in the main chains and $\alpha$-1longrightarrow2, $\alpha$-1longrightarrow3, or $\alpha$-1longrightarrow4 branch glycosidic linkages, depending on the specificity of the particular dextransucrase. Glucansucrases that catalyze glucans synthesis from sucrose. When other carbohydrates, in addition to sucrose, are present in the enzyme digest, the enzyme transfers glucose to the carbohydrate acceptors in the secondary reaction that diverts some of the glucose from incorporation into glucan. Many carbohydrate acceptors have been recognized and the products that result are dependent on the particular enzyme and the structure of the particular acceptor. Because of these unique catalytic characteristics, various dextransucrases have many important industrial and medical uses. To improve the understanding of their action mode and extend their applications, this study describes mechanism of glucan synthesis and potential industrial uses of dextransucrases, and our recent findings on the structural, functional organization and directed evolution of the glucansucrases to offer for designing glucansucrases with improved properties.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.2
• /
• pp.171-180
• /
• 2017

Objective: This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR) silage. Methods: The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR) or Leymus chinensis hay (LTMR), corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. Results: Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens), B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens) in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. Conclusion: The microbial amylase contributes to starch hydrolysis during the ensiling process in both TMR silages, whereas the microbial hemicellulase participates in the hemicellulose degradation only at the early stage of ensiling.

• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.12
• /
• pp.1725-1731
• /
• 2002

The distribution and activities of hydrolytic enzymes (cellulolyti, hemicellulolytic,pectinolytic and others) in the rumen compartments of Hereford bulls fed 100% alfalfa hay based diets were evaluated. The alfalfa proportion in the diet was gradually increased for two weeks. Whole rumen contents were processed into four fractions: Rumen contents including both the liquid and solid fractions were homogenized and centrifuged, and the supernatant was assayed for enzymes located in whole rumen contents (WRE); rumen contents were centrifuged and the supernatant was assayed for enzymes located in rumen fluids (RFE); feed particles in rumen contents were separated manually, washed with buffer, resuspended in an equal volume of buffer, homogenized and centrifuged and supernatant was assayed for enzymes associated with feed particles (FAE); and rumen microbial cell fraction was separated by centrifugation, suspended in an equal volume of buffer, sonicated and centrifuged, and the supernatant was assayed for enzymes bound with microbial cells (CBE). It was found that polysaccharide-degrading proteins such as $\beta$-1,4-D-endoglucanase, $\beta$-1,4-D-exoglucanase, xylanase and pectinase enzymes were located mainly with the cell bound (CBE) fraction. However, $\beta$-D-glucosidase, $\beta$-D-fucosidase, acetylesterase, and $\alpha$-L-arabinofuranosidase were located in the rumen fluids (RFE) fraction. Protease activity distributions were 37.7, 22.1 and 40.2%, and amylase activity distributions were 51.6, 18.2 and 30.2% for the RFE, FAE and CBE fractions, respectively. These results indicated that protease is located mainly in rumen fluid and with microbial cells, whereas amylase was located mainly in the rumen fluid.

• Journal of Microbiology and Biotechnology
• /
• v.14 no.5
• /
• pp.1075-1080
• /
• 2004

Dextransucrase (DSRB742) from Leuconostoc mesenteroides NRRL B-742CB is a glucosyltransferase that catalyzes the synthesis of dextran using sucrose, or the synthesis of oligosaccharides when acceptor molecules, like maltose, are present. The DSRB742 gene (dsrB742) was cloned and the properties were characterized. In order to identify critical amino acid residues, the DSRB742 amino acid sequence was aligned with glucosyltransferase sequences, and three amino acid residues reported as sucrose binding amino acids in Streptococcus glucosyltransferases were selected for site-directed mutagenesis experiments. Asp-533, Asp-536, and His-643 were independently replaced with Ala or Asn. D533A and D536A dextransucrases showed reduced dextran synthesis activities, 2.3% and 40.8% of DSRB742 dextransucrase, respectively, and D533N, D536N, H643A, end H643N dextransucrases showed complete suppression of dextran synthesis activities altogether. Additionally, D536N dextransucrase showed complete suppression of oligosaccharide synthesis activities. However, modifications at Asp-533 or at His-643 retained acceptor reaction activities in the range of 8.4% to 21.3% of DSRB742 acceptor reaction activity. Thus at least two carboxyl groups of Asp-533 and Asp-536, and His-643 as a proton donor, are essential for the catalysis process.

• Journal of Applied Biological Chemistry
• /
• v.43 no.4
• /
• pp.207-210
• /
• 2000

As a typical example of the screening for a microbial biocatalyst from nature, isolation of nitrilesynthesizing microorganisms, characterization of a new enzyme aldoxime dehydratase, and its function in the aldoxime-nitrile pathway are introduced. Catalytic properties of some of our enzymes were improved through a direct evolutionary approach.