• Journal of Soil and Groundwater Environment
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• v.26 no.5
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• pp.49-59
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• 2021

The effects of green manure crops, hairy vetch and sesban, supplemented with HS (humic substance) on biological soil health indicators was studied in a pot containing two kinds of reclaimed soil previously contaminated with petroleum hydrocarbons; a soil remediated by land-farming (DDC) and another soil by low-temperature thermal desorption (YJ). Treatments include no plant (C), plants only (H), and plants+2% HS (PH), which were evaluated in a pot containing respective soil. Biological indicators include microbial community analysis as well as soil enzyme activities of dehydrogenase, 𝛽-glucosidase, N-acetyl-𝛽-D-glucosaminidase (NAG), acid/alkaline phosphatase, arylsulfatase, and urease. Results showed an increase of enzyme activities in pot soils with plants and even greater in soils with plants+HS. The enzyme activities of DDC soil with plants (DDC_P) and with plants+HS (DDC_PH) increased 1.6 and 3.9 times on average, respectively than those in the control. The enzyme activities YJ soil with plants (YJ_P) and with plant+HS (YJ_PH) increased 1.8 and 3.8 times on average, respectively than those in the control. According to microbial community analysis, the relative abundance of nitrogen-fixing bacteria in DDC and YJ soil was increased from 1.5% to 7% and from 0 to 5%, respectively, after planting hairy vetch and sesban. This study showed that mixed planting of green manure crops with a supplement of humic substance is highly effective for the restoration of biological health indicators of reclaimed soils.

• Food Science and Biotechnology
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• v.17 no.5
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• pp.904-911
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• 2008

Covalent cross-links between a number of proteins and peptides explain why transglutaminase may be widely used by food processing industries. The objective of this work was optimization of the fermentation process to produce transglutaminase from a new microbial source, the Streptomyces sp. P20. The strategy adopted to modify the usual literature media was: (1) fractional factorial design (FFD) to elucidate the key medium ingredients, (2) central composite design (CCD) to optimise the concentration of the key components. Optimization of the medium resulted in not only an 86% increase in microbial transglutaminase activity as compared to the media cited in the literature, but also a reduction in the production cost. Optimal fermentation conditions - namely temperature and agitation rate - were also studied, using CCD methodology. Usual conditions of $30^{\circ}C$ and 100 rpm were within the optimal area. All other parameters for enzyme production were experimentally proven to be optimum fermentation conditions.

• Research in Plant Disease
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• v.27 no.4
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• pp.155-163
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• 2021

Dried red peppers are frequently contaminated with mycotoxins during storage. To determine the effect of storage environments on fungal occurrence and subsequent mycotoxin accumulation in dried red peppers, we monitored red pepper powder and whole fruit samples for fungal occurrence under various temperatures and relative humidity (RH) conditions during 340 days. Fungal occurrences fluctuated in both pepper forms throughout the storage but they were higher in pepper powder than whole one, higher under low temperatures (-20℃, 0℃, or 4℃) than others (10℃, 25℃, or 30℃), and higher under RH 93% than RH 51% and 69% in both peppers. The samples exhibiting high fungal occurrences were associated mainly with dominant species such as Aspergillussydowii, Penicillium solitum, P. roqueforti, P. polonicum, or P. chrysogenum. Mycotoxigenic species, including A. flavus, A. ochraceus, A. westerdijkiae, A. tubingensis, and P. citrinum, were also detected throughout the samples. Although mycotoxins were not detected in the samples, mycotoxigenic potential of A. flavus, A. ochraceus, and A. westerdijkiae isolates were confirmed. These results show that low temperatures (-20℃, 0℃, or 4℃) and/or high surrounding RH (>93%) are not safe environments for storage of dried red peppers as fungal growth can occur under these conditions.

• Food Science and Preservation
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• v.30 no.2
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• pp.287-299
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• 2023

In vitro luwak coffee was produced using enzyme­microbial complexes. The coffee quality of non-fermented coffee beans (NFC) and fermented coffee beans (FC) was compared. The total free amino acid content was higher in FC than in NFC. The levels of glutamic acid and γ-amino-n-butyric acid in NFC were higher than those in FC; however, the contents of essential amino acids, such as lysine, leucine, and valine, in FC were higher than in NFC. During fermentation, the sucrose content decreased, whereas the fructose and glucose contents increased (p<0.001). The chromaticity of the coffee extract showed higher lightness (L), redness (a), and yellowness (b) values in FC than those in NFC. The caffeine content was significantly lower in FC (696.94±0.04 ㎍/mL) compared to that in NFC (1,130.22±1.55 ㎍/mL) (p<0.001). Conversely, the polyphenol and chlorogenic acid contents were significantly higher in NFC than in FC (p<0.001). Electronic nose analysis indicated considerable differences between the volatile aromatic components in NFC and FC. Sensory scores were significantly higher for FC than those for NFC. Therefore, the fermentation of coffee beans using enzyme­microbial complexes altered the chemical components, which promoted the Maillard reaction during the coffee bean roasting process. These results suggest the possibility of producing in vitro luwak coffee with better flavor and lower caffeine content.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1547-1556
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• 2009

A novel thermostable $\alpha$-glucosidase (TtGluA) from Thermoanaerobacter tengcongensis MB4 was successfully expressed in E. coli and characterized. The TtgluA gene contained 2,253 bp, which encodes 750 amino acids. The native TtGluA was a trimer with monomer molecular mass of 89 kDa shown by SDS-PAGE. The purified recombinant enzyme showed hydrolytic activity on maltooligosaccharides, p-nitrophenyl-$\alpha$-D-glucopyranide, and dextrin with an exotype cleavage manner. TtGluA showed preference for short-chain maltooligosaccharides and the highest specific activity for maltose of 3.26 units/mg. Maximal activity was observed at $60^{\circ}C$ and pH 5.5. The half-life was 2 h at $60^{\circ}C$. The enzyme showed good tolerance to urea and SDS but was inhibited by Tris. When maltose with the concentration over 50 mM was used as substrate, TtGluA was also capable of catalyzing transglycosylation to produce $\alpha$-1,4-linked maltotriose and $\alpha$-1,6-linked isomaltooligosaccharides. More importantly, TtGluA showed exclusive regiospecificity with high yield to produce $\alpha$-1,6-linked isomaltooligosaccharides when the reaction time extended to more than 10 h.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.254-264
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• 2010

To obtain an efficient natural lignocellulolytic complex enzyme, we screened an efficient lignocellulose-degrading composite microbial system (XDC-2) from composted agricultural and animal wastes amended soil following a long-term directed acclimation. Not only could the XDC-2 degrade natural lignocelluloses, but it could also secrete extracellular xylanase efficiently in liquid culture under static conditions at room temperature. The XDC-2 degraded rice straw by 60.3% after fermentation for 15 days. Hemicelluloses were decomposed effectively, whereas the extracellular xylanase activity was dominant with an activity of 8.357 U/ml on day 6 of the fermentation period. The extracellular crude enzyme noticeably hydrolyzed natural lignocelluloses. The optimum temperature and pH for the xylanase activity were $40^{\circ}C$ and 6.0. However, the xylanase was activated in a wide pH range of 3.0-10.0, and retained more than 80% of its activity at $25-35^{\circ}C$ and pH 5.0-8.0 after three days of incubation in liquid culture under static conditions. PCR-DGGE analysis of successive subcultures indicated that the XDC-2 was structurally stable over long-term restricted and directed cultivation. Analysis of the 168 rRNA gene clone library showed that the XDC-2 was mainly composed of mesophilic bacteria related to the genera Clostridium, Bacteroides, Alcaligenes, Pseudomonas, etc. Our results offer a new approach to exploring efficient lignocellulolytic enzymes by constructing a high-performance composite microbial system with synergistic complex enzymes.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.342-345
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• 2001

A bacterial strain PH-4, which produces an enzyme catalyzing the degradation of crosslinked poly(${\gamma}$-glutamic acid) hydrogels, was isolated and identified as a Flavobacterium sp. The enzyme was obtained by the sonication of the bacterial cells preincubated in a Bouillon medium with shaking, without adding of poly(${\gamma}$-glutamic acid) as an inducer. The products of the hydrogel degraded by the crude enzyme agreed closely with the depolymerized materials in SDS-polyacrylamide gel electrophoresis using methylene blue staining, and with a glutamic acid monomer on thin-layer chromatography, thereby suggesting that strain PH-4 produced a kind of exohydrolase.

• The Korean Journal of Food And Nutrition
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• v.13 no.2
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• pp.176-180
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• 2000

Microbial protease has been interesting due to the biological roles in the producing microorganism. A thermophilic Actinomyces produing protease was isolated from soil. The optimal medium composition and culture conditions for maximum protease production was as follows 0.5% soluble starch, 0.5% yeast extract. 0.1% K2HPO4, 0.05% CaCl2, initial pH 8.0 at 50$^{\circ}C$ for 48hours. The protease was purified by the procedure of ammonium sulfate precipitation, anion exchange chromatography(LC), DEAE high performance liquid chromatography and GPC HPLC. The purification fold of the purified enzyme was increased about 22.6. The optimal pH and temperature for reaction of the purified enzyme were 7.5 and 60$^{\circ}C$. The purified enzyme was stable for the pH range from 6.0 to 8.5, but was unstable when treated at 80$^{\circ}C$ for 10 minutes. The activity of the enzyme was inhibited by Ag+ and Cu2+.

• Preventive Nutrition and Food Science
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• v.3 no.2
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• pp.128-132
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• 1998

7S globulin has been isolate from the defatted soybean meal(glycine max,. merill) by Sepharose-6B column chromatography and CM-Sephadex column chromatography. Coagulum of 7S globulin formed at a temperature of $65^{\circ}C$ in microbial enzyme treatment. In order to characterize the structure of the coagulum, three kinds of coagulum (enzyme-, calcium-and acid-induced coagulum)were compared througth the Scanning Electron Microscope(SEM). The network structure was found to be of two levels. First, there was an appearance on the molecular level in the form of strands. Second, there was a denser network with a fine structure.

• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.887-892
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• 2015

Uricase is an important microbial enzyme that can be used in the clinical treatment of gout, hyperuricemia, and tumor lysis syndrome. A total of 127 clinical isolates of Pseudomonas aeruginosa were tested for uricase production. A Pseudomonas strain named Ps43 showed the highest level of native uricase enzyme expression. The open reading frame of the uricase enzyme was amplified from Ps43 and cloned into the expression vector pRSET-B. Uricase was expressed using E. coli BL21 (DE3). The ORF was sequenced and assigned GenBank Accession No. KJ718888. The nucleotide sequence analysis was identical to the coding sequence of uricase gene puuDof P. aeruginosa PAO1. We report the successful expression of P. aeruginosa uricase in Escherichia coli. E. coli showed an induced protein with a molecular mass of about 58 kDa that was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. We also established efficient protein purification using the Ni-Sepharose column with activity of the purified enzyme of 2.16 IU and a 2-fold increase in the specific activity of the pure enzyme compared with the crude enzyme.