• Journal of Animal Science and Technology
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• v.49 no.5
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• pp.667-676
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• 2007

This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.

• Microbiology and Biotechnology Letters
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• v.35 no.3
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• pp.255-260
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• 2007

This study was conducted to isolate and identify bacteria producing xylanase and cellulase from spent mushroom substrates and to determine the optimal medium conditions for their growth. Bacteria showing high xylanase and carboxymethyl cellulase activities and low protease and amylase activities were strain 201-3 and strain 206-3. Strain 201-3 was identified as Enterobacter ludwigii and named Ent. ludwigii KU201-3. 206-3 was identified as Bacillus cereus and named B. cereus KU206-3. The optimal medium condition of Ent. ludwigii KU201-3 was obtained when 1%(w/v) of soybean meal and 3%(w/v) of sucrose were used as nitrogen and carbon source, respectively. That of B. cereus KU206-3 was obtained when 3%(w/v) of soybean meal and 1%(w/v) of molasses were used as nitrogen and carbon sources, respectively.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.37 no.1 s.109
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• pp.25-37
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• 2005

This study was carried out to examine the optimal cultural condition in enzyme activities of CMCase, FPase and xylanase in selected strains which secret extracellular enzymes for using deinking agent to old newsprint. The results of this study were as follow: The production of enzyme by Bacillus pumilus I was maximal as grown on the medium, containing of rice bran+xylan $2.0\%$, peptone $0.8\%,\;K_2HPO_4\;0.1\%\;and\;CaCl_2\;0.06\%$ at pH 8.0 and $28^{\circ}C$ for 72 hours. Optimal cultural condition of B. subtilis I was avicel+xylan $3.5\%,\;urea\;0.4\%,\;K_3PO_4\;0.1\%\;and\;CaCl_2\;0.015\%$ at pH 9.0 and $28^{\circ}C$ for 36 hours. The maximal enzyme production was observed in the medium, containing of avicel+xylan $3.5\%,\;urea\;1.6\%\;and\; K_2HPO_4\;0.125\%$ with pH 9.0 when B. pumilus II was cultured at $28^{\circ}C$ for 60 hours. The production of enzyme by B. subtilis IT was maximal as grown on the medium, containing of xylan $2.0\%,\;yeast\; extract\;0.6\%,\;K_2HPO_4\;0.1\%\;and\;ZnSO_4\;0.04\%$ at pH 8.0 and $34^{\circ}C$ for 36 hours. The activities of FPase and xylanase in tested 4 strains were not much different with Thermomonospora fusca.

• Applied Biological Chemistry
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• v.45 no.2
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• pp.71-76
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• 2002

Several antagonistic bacteria, SD-1, 4, 10, 11, 14, 15, and 16, which have strong CMCase and amylase activities, were isolated from the fermented mushroom media. Among them, SD-1, 10, 11, and 15 have strong antibacterial activities against the mushroom pathogenic bacteria Pseudomonas sp., and SD-1, 10, 11, 14, and 16 have strong antifungal activities against the mushroom pathogenic fungi, Trichoderma sp. SD-14, 15, and 16 did not inhibit the growth of mushroom Pleurotus eryngii ASI-2302, and Pleurotus ostreatus ASI-2042 and ASI-2180. When the culture broth mixture of the seven bacterial strains was applied to the mushroom media, the growths of pathogens, Pseudomonas sp. and Trichoderma sp., were inhibited.

• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.588-596
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• 2018

An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.