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( Zhuang Yao ),( Xiaoming Liu ),( Jae Min Shim ),( Kang Wook Lee ),( Hyun-jin Kim ),( Jeong Hwan Kim ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.1
Nine bacilli with fibrinolytic activities were isolated from doenjang, a traditional Korean fermented soy food. Among them, RSB34 showed the strongest activity and was identified as Bacillus amyloliquefaciens by 16S rRNA and recA gene sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity (83.23 mU/μl) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when the culture supernatant was analyzed by SDSPAGE and 27 and 42 kDa bands by fibrin zymography. The gene encoding the 27 kDa fibrinolytic enzyme AprE34 was cloned by PCR. BLAST analyses confirmed that the gene was a homolog to genes encoding AprE-type proteases. aprE34 was overexpressed in Escherichia coli BL21(DE3) using pET26b(+). Recombinant AprE34 was purified and examined for its properties. The K<sub>m</sub> and V<sub>max</sub> values of recombinant AprE34 were 0.131 ± 0.026 mM and 16.551 ± 0.316 μM/l/min, respectively, when measured using an artificial substrate, Nsuccinyl- ala-ala-pro-phe-p-nitroanilide. aprE34 was overexpressed in B. subtilis WB600 using pHY300PLK. B. subtilis transformants harboring pHYRSB34 (pHY300PLK with aprE34) showed higher fibrinolytic activity than B. amyloliquefaciens RSB34.
Min Tan,Zhen Shang,Chenbo Xie,Hui Ma,Qian Deng,Xiaomin Tian,Peng Zhuang,Zhanye Zhang,Yingjian Wang 한국광학회 2018 Current Optics and Photonics Vol.2 No.1
To measure atmospheric temperature, water vapor, and aerosol simultaneously, an efficient multi-function Raman lidar using an ultraviolet-wavelength laser has been developed. A high-performance spectroscopic box that utilizes multicavity interference filters, mounted sequentially at small angles of incidence, is used to separate the lidar return signals at different wavelengths, and to extract the signals with high efficiency. The external experiments are carried out for simultaneous detection of atmospheric temperature, water vapor, and aerosol extinction coefficient in Beijing, under clear and hazy weather conditions. The vertical profiles of temperature, water vapor, and aerosol extinction coefficient are analyzed. The results show that for an integration time of 5 min and laser energy of 200 mJ, the mean deviation between measurements obtained by lidar and radiosonde is small, and the overall trend is similar. The statistical temperature error for nighttime is below 1 K up to a height of 6.2 km under clear weather conditions, and up to a height of 2.5 km under slightly hazy weather conditions, with 5 min of observation time. An effective range for simultaneous detection of temperature and water vapor of up to 10 km is achieved. The temperature-inversion layer is found in the low troposphere. Continuous observations verify the reliability of Raman lidar to achieve real-time measurement of atmospheric parameters in the troposphere.
조민정 ( Min Jeong Cho ),심재민 ( Jae Min Shim ),이재용 ( Jae Yong Lee ),이강욱 ( Kang Wook Lee ),야오좡 ( Zhuang Yao ),류샤오밍 ( Xiaoming Liu ),김정환 ( Jeong Hwan Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2016 한국미생물·생명공학회지 Vol.44 No.2
대두에 B. amyloliquefaciens 2 균주들과 P. farinose SY80, 그리고 R. oryzae 총 4 균주를 접종한 메주를 만들었다(메주 1). 대조구로 B. subtilis KACC16450과 A. oryzae를 접종한 메주(메주 2)와 볏짚을 균원시료로 접종한 메주(메주 3)들을 제조하여 총 56일 발효시켰다. 발효 기간중 메주 1과 2의 pH와 적정산도는 메주 3보다 각각 높고 그리고 낮았다. 메주의 혈전용해능은 메주 1이 가장 높았다. 발효 기간중 모든 메주에서 B. cereus는 검출되지 않았으나 메주 2와 3에서 여러 모양과 색을 띈 잡균들이 검출되었다. 메주 2에서 histamine이 그리고 메주 3에서 tyramine이 저 농도로 검출되었으나 메주 1에서는 둘 다 검출되지 않았다. B. amyloliquefaciens EMD17이나 MJ1-4와 같은 항균력 균주들을 접종하여 메주를 제조하면 미생물학적으로 보다 안전한 장류제품들 생산이 가능할 것이다. A test meju (meju 1) was prepared by inoculating two strains of Bacillus amyloliquefaciens (EMD17, MJ1-4), Pichia farinosa SY80, and Rhizopus oryzae into cooked soybeans. A control (meju 2) was prepared by inoculating Bacillus subtilis KACC16450 and Aspergillus oryzae. Another control (meju 3) was prepared using rice straw as the source of microorganisms. Three different mejus were fermented for 56 days outdoors. Meju 1 and meju 2 showed higher pH values than meju 3, whereas meju 3 showed higher titratable acidity than meju 1 and meju 2. Meju 1 showed the highest fibrinolytic activity. Bacillus cereus was not detected in any mejus, but various microorganisms were detected in meju 2 and 3. Histamine was detected in meju 2 and tyramine in meju 3 at lower concentration, which were not detected in meju 1. It was concluded that microbially safe, fermented soybean products could be produced from meju fermented with starters such as B. amyloliquefaciens EMD17 and B. amyloliquefaciens MJ1-4.
( Zhuang Ding ),( Tao Tao ),( Lili Wang ),( Yanna Zhao ),( Huiming Huang ),( Demeng Zhang ),( Min Liu ),( Zhengping Wang ),( Jun Han ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.5
Endophytic fungi are an important component of plant microbiota, and have the excellent capacity for producing a broad variety of bioactive metabolites. These bioactive metabolites not only affect the survival of the host plant, but also provide valuable lead compounds for novel drug discovery. In this study, forty-two endophytic filamentous fungi were isolated from Ficus elastica leaves, and further identified as seven individual taxa by ITS-rDNA sequencing. The antimicrobial activity of these endophytic fungi was evaluated against five pathogenic microorganisms. Two strains, Fes1711 (Penicillium funiculosum) and Fes1712 (Trichoderma harzianum), displayed broad-spectrum bioactivities. Our following study emphasizes the isolation, identification and bioactivity testing of chemical metabolites produced by T. harzianum Fes1712. Two new isocoumarin derivatives (1 and 2), together with three known compounds (3-5) were isolated, and their structures were elucidated using NMR and MS. Compounds 1 and 2 exhibited inhibitory activity against Escherichia coli. Our findings reveal that endophytic fungi from the rubber tree F. elastica leaves exhibit unique characteristics and are potential producers of novel natural bioactive products.
DNA Shuffling of aprE Genes to Increase Fibrinolytic Activity and Thermostability
Yao, Zhuang,Jeon, Hye Sung,Yoo, Ji Yeon,Kang, Yun Ji,Kim, Min Jae,Kim, Tae Jin,Kim, Jeong Hwan The Korean Society for Microbiology and Biotechnol 2022 Journal of microbiology and biotechnology Vol.32 No.6
Four aprE genes encoding alkaline serine proteases from B. subtilis strains were used as template genes for family gene shuffling. Shuffled genes obtained by DNase I digestion followed by consecutive primerless and regular PCR reactions were ligated with pHY300PLK, an E. coli-Bacillus shuttle vector. The ligation mixture was introduced into B. subtilis WB600 and one transformant (FSM4) showed higher fibrinolytic activity. DNA sequencing confirmed that the shuffled gene (aprEFSM4) consisted of DNA mostly originated from either aprEJS2 or aprE176 in addition to some DNA from either aprE3-5 or aprESJ4. Mature AprEFSM4 (275 amino acids) was different from mature AprEJS2 in 4 amino acids and mature AprE176 in 2 amino acids. aprEFSM4 was overexpressed in E. coli BL21 (DE3) by using pET26b(+) and recombinant AprEFSM4 was purified. The optimal temperature and pH of AprEFSM4 were similar to those of parental enzymes. However, AprEFM4 showed better thermostability and fibrinogen hydrolytic activity than the parental enzymes. The results indicated that DNA shuffling could be used to improve fibrinolytic enzymes from Bacillus sp. for industrial applications.
( Xuelong Zhuang ),( Min Chang Shin ),( Byeong Jun Jeong ),( Seung Hwan Lee ),( Jung Hoon Park ) 한국화학공학회 2021 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.59 No.2
Recently, hybrid coal research is underway to upgrade low-grade coal. The hybrid coal is made by mixing low-grade coal with bioliquids such as molasses, sugar cane, and lignin. In the case of lignin used here, a large amount of lignin is included in the wastewater of the papermaking process, and thus, research on hybrid coal production using the same is attracting attention. However, since a large amount of metal ions are contained in the lignin wastewater from the papermaking process, substances that corrode the generator are generated during combustion, and the amount of fly ash is increased. To solve this problem, it is essential to remove metal ions in the lignin wastewater. In this study, metal ions were removed by ion exchange with a alumina hollow fiber membrane coated with K-Phillipsite (K-PHI) zeolite. The alumina hollow fiber membrane used as the support was prepared by the nonsolvent induced phase separation (NIPS) method, and K-PHI seeds were prepared by hydrothermal synthesis. The prepared K-PHI seed was seeded on the surface of the support and coated by secondary growth hydrothermal synthesis. The characteristic of prepared coating membrane was analyzed by Scanning Electron Microscope (SEM), X-Ray Diffraction (XRD), Energy Dispersive Spectroscopy (EDX), and the concentration of metal ions before and after ion exchange was measured by Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES). The extraction amount of K<sup>+</sup> is 86 mg/kg, and the extraction amount of Na<sup>+</sup> is 54.9 mg/kg. Therefore, K-PHI zeolite membrane has the potential to remove potassium and sodium ions from the solution and can be used in acidic lignin wastewater.