Endophytic Bacillus subtilis MJMP2 from Kimchi inhibits Xanthomonas oryzae pv. oryzae, the pathogen of Rice bacterial blight disease

( Jinhua Cheng ),( Kumar Sagar Jaiswal ),( Seung Hwan Yang ),( Joo Won Suh ) 한국응용생명화학회 2016 Journal of Applied Biological Chemistry (J. Appl. Vol.59 No.2

An endophytic bacterial strain was isolated from kimchi, a Korean traditional fermented Brassica campestris and identified as Bacillus subtilis MJMP2 based on the 16S rRNA sequence. This strain showed strong antagonistic activity against Xanthomonas oryzae pv. oryzae (Xoo) KACC10331, the pathogen of bacterial rice blight disease, as well as activity against some other rice phytopathogenic fungi. The active compound was purified through size-exclusion chromatography and preparative High-performance liquid chromatography. The molecular weight was determined as m/z 1043 by mass spectroscopy, which is identical to that of iturin A. Furthermore, a crude extract from the culture supernatant of Bacillus subtilis MJMP2 showed inhibitory activity against rice blight disease in both a rice leaf explant assay and a pot assay. The crude extract also enhanced the length of roots of Arabidopsis thaliana. These results suggest that the strain Bacillus subtilis MJMP2 could be used as a biological agent to control rice blight disease.

Azalomycin F Complex Is an Antifungal Substance Produced by Streptomyces malaysiensis MJM1968 Isolated from Agricultural Soil

Cheng, Jinhua,Yang, Seung-Hwan,Palaniyandi, Sasikumar Arunachalam,Han, Jung-Sun,Yoon, Tae-Mi,Kim, Tae-Jong,Suh, Joo-Won The Korean Society for Applied Biological Chemistr 2010 Applied Biological Chemistry (Appl Biol Chem) Vol.53 No.5

MJM1968, an actinomycete showing potent antifungal activity, was isolated and identified by 16S rDNA sequence analysis as Streptomyces malaysiensis. MJM1968 showed strong antifungal activity on phytopathogenic fungi such as Fusarium oxysporum, Rhizoctonia solani, Cladosporium cladosporioides, Fusarium chlamydosporum, Colletotrichum gloeosporioides KACC 40693, Alternaria mali KACC 40026, and Pestalotia spp. KACC 40501, in vitro. An antifungal compound was isolated from culture filtrate of strain MJM1968 by a series of chromatographic methods. Treatment of agricultural soil with strain MJM1968 mycelia reduced the native fungal population by more than 60% after 14 days. Treatment of soil with 10 mg/mL partially purified extract lowered native fungal density by more than 80% after 14 days. The compound, which exhibited antifungal activity in a broad range of pH and temperature, was identified as azalomycin F complex by $^1H$ and $^{13}C$ NMRs. This is the first report on the isolation of azalomycin F complex from S. malaysiensis, demonstrating a broad-spectrum suppression of fungal pathogens in agricultural soil.

Template Synthesis of Nitrogen-Doped Short Tubular Carbons with Big Inner Diameter and their Application in Electrochemical Sensing

Cheng, Rui,Zou, Qiong,Zhang, Xiaohua,Xiao, Chunhui,Sun, Longfei,Chen, Jinhua Korean Chemical Society 2014 Bulletin of the Korean Chemical Society Vol.35 No.8

Nitrogen-doped short tubular carbons (N-STCs) with big inner diameter have been successfully synthesized via carbonization of polydopamine (PDA) wrapped halloysite nanotubes (HNTs). The obtained N-STCs have average length of $0.3{\mu}m$ with big inner diameter (50 nm), thin wall (2-3 nm) and large surface area ($776m^2g^{-1}$), and show excellent electrochemical properties. As an example in electrochemical applications, N-STCs were used to electrochemically detect hydrogen peroxide ($H_2O_2$) and glucose. The results showed that the N-STCs modified glassy carbon (N-STCs/GC) electrode had much better analytical performance (lower detection limit and wider linear range) compared to the acid-treated carbon nanotubes (AO-CNTs) based GC electrode. The unique structure endows N-STCs the enhanced electrochemical performance and promising applications in electrochemical sensing.

Template Synthesis of Nitrogen-Doped Short Tubular Carbons with Big Inner Diameter and their Application in Electrochemical Sensing

Rui Cheng,Qiong Zou,Xiaohua Zhang,Chunhui Xiao,Longfei Sun,Jinhua Chen 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.8

Nitrogen-doped short tubular carbons (N-STCs) with big inner diameter have been successfully synthesized via carbonization of polydopamine (PDA) wrapped halloysite nanotubes (HNTs). The obtained N-STCs have average length of 0.3 μm with big inner diameter (50 nm), thin wall (2-3 nm) and large surface area (776 m2 g−1), and show excellent electrochemical properties. As an example in electrochemical applications, N-STCs were used to electrochemically detect hydrogen peroxide (H2O2) and glucose. The results showed that the N-STCs modified glassy carbon (N-STCs/GC) electrode had much better analytical performance (lower detection limit and wider linear range) compared to the acid-treated carbon nanotubes (AO-CNTs) based GC electrode. The unique structure endows N-STCs the enhanced electrochemical performance and promising applications in electrochemical sensing.

Differential anticancer effect of fermented squid jeotgal due to varying concentrations of soymilk additive

( Fahima Akther ),( Jinhua Cheng ),( Seung Hwan Yang ),( Gyuhwa Chung ) 한국응용생명화학회 2017 Journal of Applied Biological Chemistry (J. Appl. Vol.60 No.2

Fermentation plays a vital role in the nutritional enrichment of food. Korea has a long tradition of adding fermented food to the daily diet and jeotgal is one of the common fermented and salted foods in Korean cuisine. In our study, we added soymilk as an additive to squid jeotgal to improve its functionality. We mixed different concentrations of soymilk (2, 5, and 10 mg/g) with squid jeotgal samples, fermented them for one week, and then tested their antioxidant and anticancer activities to compare with those of squid jeotgal samples without soymilk additive. To investigate the anticancer characteristics, glutathione-S-transferase (GST)-pi enzyme assay was used. To test the antioxidant activities, various assays were performed, including 2,2-diphenyl-1-picryl hydrazyl free radical scavenging activity, 2,2-azino-bis(3-ethyl-benzothiazoline- 6-sulfonic acid) diammonium saltradical cation scavenging assay, and reducing power assay. Samples fermented with a small amount of soymilk showed excellent anticancer activity. The addition of only 2 mg/g of soymilk to squid jeotgal inhibited the activity of GST-pi by almost 50% when compared with the sample with no addition. Moreover, no undesirable bitterness or astringency was noticed. Our results could help to improve the current food status of squid jeotgal and it could be used to reduce the risk of chronic disease along with its basic nutritional function.

<i>S</i>-Adenosyl-<small>L</small>-methionine Activates Actinorhodin Biosynthesis by Increasing Autophosphorylation of the Ser/Thr Protein Kinase AfsK in <i>Streptomyces coelicolor</i> A3(2)

JIN, Ying-Yu,CHENG, Jinhua,YANG, Seung Hwan,MENG, Lingzhu,PALANIYANDI, Sasikumar Arunachalam,ZHAO, Xin-Qing,SUH, Joo-Won Japan Society for Bioscience, Biotechnology, and A 2011 Bioscience, biotechnology, and biochemistry Vol.75 No.5

<P><I>S</I>-Adenosyl-<SMALL>L</SMALL>-methionine (SAM) is one of the major methyl donors in all living organisms. The exogenous treatment with SAM leads to increased actinorhodin production in <I>Streptomyces coelicolor</I> A3(2). In this study, mutants from different stages of the AfsK-AfsR signal transduction cascade were used to test the possible target of SAM. SAM had no significant effect on actinorhodin production in <I>afsK</I>, <I>afsR</I>, <I>afsS</I>, or <I>act</I>II-open reading frame 4 (ORF4) mutant. This confirms that <I>afsK</I> plays a critical role in delivering the signal generated by exogenous SAM. The <I>afsK-pHJL-KN</I> mutant did not respond to SAM, suggesting the involvement of the C-terminal of AfsK in binding with SAM. SAM increased the <I>in vitro</I> autophosphorylation of kinase AfsK in a dose-dependent manner, and also abolished the effect of decreased actinorhodin production by a Ser/Thr kinase inhibitor, K252a. In sum, our results suggest that SAM activates actinorhodin biosynthesis in <I>S. coelicolor</I> M130 by increasing the phosphorylation of protein kinase AfsK.</P>

Original Article: Bioactive Materials : Screening the Antibacterial Activities of Streptomyces Extracts against Phytopathogens Xanthomonas oryzae pathovar oryzae, Xanthomonas campestris pathovar vesicatoria, and Pectobacterium carotovorum pathovar carotov

( Seung Hwan Kim ),( Jinhua Cheng ),( Seung Hwan Yang ),( Joo Won Suh ),( Eun Sung Song ),( Lin Woo Kang ),( Jeong Gu Kim ) 한국응용생명화학회 2015 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.3

Xanthomonas oryzae pv. oryzae (Xoo), X. campestris pv. vesicatoria (Xcv), and Pectobacterium carotovorum pv. carotovorum (Pcc) are the causative agents of bacterial blight in rice, bacterial spot in pepper, and bacterial soft rot in carrot and cabbage, respectively. To isolate novel microbial extracts with antimicrobial activities against these bacteria, approximately 5,300 different Streptomyces extracts were prepared and tested. Microbial cultures from various Streptomyces strains isolated from the Jeju Island, Baekam, Mankyoung river, Jiri mountain etc. in Korea were extracted into three different factions -secreted hydrophobic, secreted hydrophilic, and mycelia- using ethyl acetate, water, and methanol. Initially, 34, 29, and 10 extracts were selected as having antibacterial activities against Xoo, Xcv, and Pcc, respectively. Extracts 1169G4, 1172E9, and 1172E10 had the highest growth inhibition activities against both Xoo and Xcv, and extracts 1151H7 and 1152H7 showed the highest growth inhibition activities against Pcc.

Antibiotic Resistance Mechanisms Inform Discovery: Identification and Characterization of a Novel <i>Amycolatopsis</i> Strain Producing Ristocetin

Truman, Andrew W.,Kwun, Min Jung,Cheng, Jinhua,Yang, Seung Hwan,Suh, Joo-Won,Hong, Hee-Jeon American Society for Microbiology 2014 Antimicrobial Agents and Chemotherapy Vol.58 No.10

<P>Discovering new antibiotics is a major scientific challenge, made increasingly urgent by the continued development of resistance in bacterial pathogens. A fundamental understanding of the mechanisms of bacterial antibiotic resistance will be vital for the future discovery or design of new, more effective antibiotics. We have exploited our intimate knowledge of the molecular mechanism of glycopeptide antibiotic resistance in the harmless bacterium <I>Streptomyces coelicolor</I> to develop a new two-step cell wall bioactivity screen, which efficiently identified a new actinomycete strain containing a previously uncharacterized glycopeptide biosynthetic gene cluster. The screen first identifies natural product extracts capable of triggering a generalized cell wall stress response and then specifically selects for glycopeptide antibacterials by assaying for the induction of glycopeptide resistance genes. In this study, we established a diverse natural product extract library from actinomycete strains isolated from locations with widely varying climates and ecologies, and we screened them using the novel two-step bioassay system. The bioassay ultimately identified a single strain harboring the previously unidentified biosynthetic gene cluster for the glycopeptide ristocetin, providing a proof of principle for the effectiveness of the screen. This is the first report of the ristocetin biosynthetic gene cluster, which is predicted to include some interesting and previously uncharacterized enzymes. By focusing on screening libraries of microbial extracts, this strategy provides the certainty that identified producer strains are competent for growth and biosynthesis of the detected glycopeptide under laboratory conditions.</P>

Amycolatopsis sp. KCTC 29142로부터 유래된 siderochelin A의 다제 내성 균주에 대한 항균활성

이동령,성금화,이성권,홍희전,송재경,양승환,서주원,Lee, Dong-Ryung,Cheng, Jinhua,Lee, Sung-Kwon,Hong, Hee-Jeon,Song, Jaekyeong,Yang, Seung Hwan,Suh, Joo-Won The Microbiological Society of Korea 2016 미생물학회지 Vol.52 No.3

본 연구에서는 신규 Amycolatopsis 균주 KCTC 29142를 분리하여 형태학적 관찰, 계통분석 및 화학분류학적 분석 등 다상 분류분석을 통해 분석하였다. KCTC 29142 균주의 에틸아세테이트추출물은 강한 항균활성을 나타났고, 활성물질은 철 이온 킬레이트 물질인 siderochelin A로 동정되었다. 본 연구에서 분리된 siderochelin A는 다제내성균인 Acinetobacter baumanii, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), 및 Escherichia coli (E. coli)에 대해 강한 활성을 보였고, 임상에서 분리된 다제내성균에 대한 MIC를 결정하였다. A novel Amycolatopsis strain KCTC 29142 was isolated and characterized based on the polyphasic taxonomic analysis including morphological observation, phylogenetic analysis, physiological and chemotaxonomic characteristics. The ethyl acetate extract of strain KCTC 29142 culture broth showed strong antibacterial activity and the active compound was identified as siderochelin A, a ferrous-ion chelating compound. In this study, siderochelin A showed good activity against multi-drug resistant pathogens, including Acinetobacter baumanii, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), and Escherichia coli (E. coli). The minimum inhibitory activity against clinical isolates was also determined.

Endosulfan으로 오염된 현장토양의 복합미생물제제를 이용한 생물학적 정화

박찬호(Chan-Ho Park),이주영(Joo-Young Lee),조민지(Min-Ji Cho),성금화(Jinhua Cheng),염규진(Kyu-Jin Yum) 대한환경공학회 2020 대한환경공학회지 Vol.42 No.11

목적: 잔류성 유기염소계 농약의 사용은 금지 되었지만 여전히 많은 농경지에서 검출되고 있다. 본 연구에서는 다양한 후보 미생물 중에서 최종 선발된 Phanerochaete chrysosporium Y-2 (KCCM-10725P) 및 Streptomyces sp. MJM14747 (KACC 81078BP) 균주를 복합미생물 형태로 현장토양에 적용하여 endosulfan 제거율을 평가하였다. 방법: 현장 시험용 부지는 대조구 및 시험구로 각각 3 m × 3 m 면적으로 설정하였다. 두 site에 35% endosulfan 유제를 살포하여 최종 농도가 약 20 ㎍/g이 되도록 하였다. 그 후 아무 처리를 하지 않은 톱밥 12 kg 및 복합미생물이 담지된 톱밥 12 ㎏을 두 site 토양에 각각 혼합하였다. 총 100일 동안의 실험 기간 중 시료는 30일 단위로 채취하였다. 채취된 토양 시료는 QuEChERS AOAC 방법에 따라 추출 및 전처리하였고 gas chromatography로 분석하였다. 결과 및 토의: 대조구 및 실험구에 endosulfan 유제를 처리한 직후의 농도는 각각 21.2 ㎍/g 및 21.9 ㎍/g으로 분석되었다. 미생물을 처리하지 않은 대조구 사이트의 endosulfan 제거율은 30일까지 약 21.9%를 나타냈고, 100일째에는 54.4%를 나타냈다. 반면, 복합미생물을 처리한 사이트의 100일째 endosulfan 제거율은 89.5%로 대조구에 비하여 1.6배 높게 나타났다. 30일째의 제거율은 56.8%였고 이후 endosulfan의 제거율이 빠르게 증가하는 경향을 보였다. Endosulfan의 주요 성분 중 α-endosulfan의 제거율이 β-endosulfan보다 높게 나타났다. 100일 경과 시 α-endosulfan과 β-endosulfan의 제거율은 각각 98.0% 및 83.5%로 확인되었다. 결론: 미생물복합제제로 선발된 Streptomyces sp. MJM14747은 100 ㎍/g 내외의 고농도 endosulfan 존재하에서도 분해능이 우수하였고, Phanerochaete chrysosporium Y-2은 endosulfan의 분해속도가 느린 β-endosulfan의 제거에 우수하였다. 복합미생물제제는 앞으로 잔류농약 문제에 직면해 있는 국내외 농경지 토양뿐만 아니라 생물학적 정화사업 전반에 응용 가능할 것으로 기대된다. Objectives : The use of risky and persistent organochlorine pesticides was prohibited, but is still detected in many agricultural lands. In this study, Phanerochaete chrysosporium Y-2 (KCCM-10725P) and Streptomyces sp. MJM14747 (KACC 81078BP), which were finally selected among various candidate microorganisms, were applied to field soil and estimated the endosulfan removal rate. Methods : The field testing sites were set to an area of 3 m × 3 m, respectively. A 35% endosulfan emulsion was sprayed on both sites so that the final concentration was about 20 ㎍/g. Thereafter, 12 ㎏ of original sawdust and 12 ㎏ of sawdust loaded with microbial consortia were mixed at each of the two sites. During the test period for a total of 100 days, samples were taken every 30 days. Soil samples were extracted according to the QuEChERS AOAC method and analyzed by gas chromatography. Results and Discussion : The concentrations immediately after endosulfan emulsion treatment at both sites were detected as 21.2 ㎍/g and 21.9 μg/g, respectively. The removal rate of endosulfan at the site without microbial consortia treatment (Control) was about 21.9% until the 30th day, and 54.4% on the 100th day. On the other hand, the endosulfan removal rate on the 100th day of the site treated with microbial consortia was 89.5%, which was 1.6 times higher than that of the control site. On the 30th day, the removal rate was 56.8%, and the rate of degradation of endosulfan increased thereafter. Among the major components of endosulfan, the removal rate of α -endosulfan was higher than that of β-endosulfan. After 100 days, the removal rates of α-endosulfan and β -endosulfan were 98.0% and 83.5%, respectively. Conclusions : Streptomyces sp. MJM14747 showed good degradation ability in the presence of endosulfan at a high concentration of around 100 ㎍/g. Moreover, Phanerochaete chrysosporium Y-2 has a good performance in the removal of β-endosulfan, which is slow degrading components of endosulfan. Microbial consortia are expected to be applied not only to domestic and foreign agricultural land but also to overall bioremediation projects.