• Title/Summary/Keyword: Biosurfactants

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미생물을 이용한 원유 회수증진법에 대한 동향연구 (Brief Review on Microbial Enhanced Oil Recovery)

  • 오경석
    • 한국응용과학기술학회지
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    • 제38권4호
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    • pp.1010-1019
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    • 2021
  • 유전에 매장된 석유는 1차, 2차, 그리고 3차에 걸쳐 회수된다. 3차 회수로 분류된 공법 중 미생물을 이용한 원유 회수증진법은 크게 in-situ와 ex-situ 공법으로 나눌 수 있다. In-situ 공법은 미생물을 저류층에 직접 투입하여 미생물의 대사활동을 통한 원유의 회수를 유도하는 공법이다. 대사물질 중 바이오 계면활성제는 저류층 잔존 유분의 유동화에 큰 역할을 한다. 한편, ex-situ 공법은 외부에서 미생물의 대사물질을 추출하여 저류층에 투입하여 원유를 회수하는 방법이다. 두 가지 공법 모두 친환경적이지만, 보다 경제적인 in-situ 공법이 선호된다. 미생물을 이용한 원유 회수증진법은 오랜기간 검토되었음에도, 현재까지도 파일럿 규모에서 여전히 평가가 진행 중이다. 본 논문에서는, in-situ 공법에 적용가능한 미생물 중 박테리아의 종 특성과 대사물질에 대해서 살펴보았다.

Identification, Characterization, and Efficacy Evaluation of Bacillus velezensis for Shot-Hole Disease Biocontrol in Flowering Cherry

  • Han, Viet-Cuong;Yu, Nan Hee;Yoon, Hyeokjun;Ahn, Neung-Ho;Son, Youn Kyoung;Lee, Byoung-Hee;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • 제38권2호
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    • pp.115-130
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    • 2022
  • Though information exists regarding the pathogenesis of the shot-hole disease (SH) in flowering cherry (FC), there has been a lack of research focusing on SH management. Therefore, here, we investigated the inhibitory activities of antagonistic bacteria against SH pathogens both in vitro and in vivo as well as their biochemical characteristics and bioactive compounds. Two biosurfactant-producing bacterial antagonists, identified as Bacillus velezensis strains JCK-1618 and JCK-1696, exhibited the best effects against the growth of both bacterial and fungal SH pathogens in vitro through their cell-free culture filtrates (CFCFs). These two strains also strongly inhibited the growth of the pathogens via the action of their antimicrobial diffusible compounds and antimicrobial volatile organic compounds (VOCs). Crude enzymes, solvent extracts, and biosurfactants of the two strains exhibited antimicrobial activities. Liquid chromatography/electrospray ionization time-of-flight mass spectrometric analysis of the partially purified active fractions revealed that the two antagonists produced three cyclic lipopeptides, including iturin A, fengycin A, and surfactin, and a polyketide, oxydifficidin. In a detached leaf assay, pre-treatment and co-treatment of FC leaves with the CFCFs led to a large reduction in the severity of the leaf spots caused by Epicoccum tobaicum and Bukholderia contaminans, respectively. In addition, the two antagonists produced indole-3-acetic acid, siderophore, and a series of hydrolytic enzymes, along with the formation of a substantial biofilm. To our knowledge, this is the first report of the antimicrobial activities of the diffusible compounds and VOCs of B. velezensis against the SH pathogens and their efficiency in the biocontrol of SH.

Statistical Optimization of Biosurfactant Production from Aspergillus niger SA1 Fermentation Process and Mathematical Modeling

  • Mansour A. Al-hazmi;Tarek A. A. Moussa;Nuha M. Alhazmi
    • Journal of Microbiology and Biotechnology
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    • 제33권9호
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    • pp.1238-1249
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    • 2023
  • In this study, we sought to investigate the production and optimization of biosurfactants by soil fungi isolated from petroleum oil-contaminated soil in Saudi Arabia. Forty-four fungal isolates were isolated from ten petroleum oil-contaminated soil samples. All isolates were identified using the internal transcribed spacer (ITS) region, and biosurfactant screening showed that thirty-nine of the isolates were positive. Aspergillus niger SA1 was the highest biosurfactant producer, demonstrating surface tension, drop collapsing, oil displacement, and an emulsification index (E24) of 35.8 mN/m, 0.55 cm, 6.7 cm, and 70%, respectively. This isolate was therefore selected for biosurfactant optimization using the Fit Group model. The biosurfactant yield was increased 1.22 times higher than in the nonoptimized medium (8.02 g/l) under conditions of pH 6, temperature 35℃, waste frying oil (5.5 g), agitation rate of 200 rpm, and an incubation period of 7 days. Model significance and fitness analysis had an RMSE score of 0.852 and a p-value of 0.0016. The biosurfactant activities were surface tension (35.8 mN/m), drop collapsing (0.7 cm), oil displacement (4.5 cm), and E24 (65.0%). The time course of biosurfactant production was a growth-associated phase. The main outputs of the mathematical model for biomass yield were Yx/s (1.18), and µmax (0.0306) for biosurfactant yield was Yp/s (1.87) and Yp/x (2.51); for waste frying oil consumption the So was 55 g/l, and Ke was 2.56. To verify the model's accuracy, percentage errors between biomass and biosurfactant yields were determined by experimental work and calculated using model equations. The average error of biomass yield was 2.68%, and the average error percentage of biosurfactant yield was 3.39%.

Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • 제20권2호
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

여러 식물병원성 진균을 억제하는 Streptomyces costaricanus HR391의 항진균능 (Antifungal activity of Streptomyces costaricanus HR391 against some plant-pathogenic fungi)

  • 김혜령;송홍규
    • 미생물학회지
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    • 제52권4호
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    • pp.437-443
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    • 2016
  • 여러 토양에서 분리한 400여 개의 방선균 균주에 대해 4가지 식물병원성 진균에 대한 항진균 활성을 조사하였으며 그 중 Streptomyces costaricanus HR391 균주는 PDB 배지에서 식물병원성 진균인 Fusarium oxysporum f. sp. raphani, F. oxysporum f. sp. niveum, F. oxysporum f. sp. lycopersici와 Rhizoctonia solani의 균사 생장을대조군과 비교하여 각각 26.5, 26.2, 21.2와 23.8% 저해하였다. S. costaricanus HR391은 항진균물질인 siderophore를 $98{\mu}M$을 생성할 뿐만 아니라 유화활성을 나타내며 막지질을 파괴할 수 있는 생물계면활성제인 rhamnolipid와 lipopeptide인 iturin A와 surfactin를 생성하였다. 또한 진균 세포막을 분해할 수 있는 chitinase와 glucanase 활성도 나타내었으며 병원균의 막을 파괴하는 AMP와 항생물질 phenazine도 분비하였다. 이외에도 식물생장 촉진활성을 갖는 zeatin, gibberellin과 indole acetic acid 같은 식물호르몬도 생성하였다. 이와 같이 항진균 활성을 나타내는 S. costaricanus HR391 균주는 다양한 종류의 항진균 물질의 상승작용과 더불어 높은 생물계면활성이 본 균주의 항진균 활성에 큰 역할을 하는 것으로 보이며 친환경적인 생물학적 항진균제로서 활용이 가능할 것으로 기대된다.

전통고추장에서 유해균 억제 및 Biogenic Amines 분해 능력을 가지는 Bacillus licheniformis SCK B11의 분리 (Antimicrobial and Biogenic Amine-Degrading Activity of Bacillus licheniformis SCK B11 Isolated from Traditionally Fermented Red Pepper Paste)

  • 김용상;정진오;조성호;정도연;엄태붕
    • 미생물학회지
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    • 제48권2호
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    • pp.163-170
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    • 2012
  • 장류의 유해균과 biogenic amines 함량을 줄이기 위해 항균작용과 biogenic amines 분해능력이 있는 한 균주를 83종의 전통장류로부터 분리하였다. 형태 및 생화학적 특성, 16S rRNA 유전자 서열 해독 결과 이 균주는 Bacillus licheniformis에 속했으며, B. licheniformis SCK B11로 명명되었다. SCK B11을 이틀간 배양한 후의 원심분리 상층액은 유해 균주들인 Enterococcus faecalis, Listeria monocytogenes, Micrococcus luteus, Pseudomonas aeruginosa, Bacillus cereus, Staphylococcus aureus, Aspergillus flavus에 대해 균 증식 저지력을 보였다. 이 균주의 항균 특성을 확인하기 위해 B. licheniformis의 항균물질로 알려진 surfactin, lichenysin, lichenicidine 합성 유전자들에 대한 PCR을 수행한 결과 모두 음성으로 나와 이 항균 물질은 기존에 보고된 물질들과는 다름을 보였다. 또한 전자 현미경 사진 결과 이 항균 물질은 유해균들의 세포막 손상을 야기하는 것이 관찰되었다. 이 균주는 각각 5.3%의 histamine과 tyramine이 함유된 삶은 콩에서 10일의 발효 기간 동안 histamine 함량의 72%, tyramine 함량의 66%를 분해하는 것으로 나타났다. SCK B11의 항균 및 biogenic amine 분해 특성을 고려할 때, 공정상 필연적으로 유해균 증식 및 biogenic amine 생성 문제를 지닌 전통 장류 발효 과정에서 이 균주는 이 문제점들을 해결할 수 있을 것으로 보인다.

Bacillus sp. TBM40-3에 의해 생성된 Biosurfactant의 유류분해 특성 (Characterization of Oil-Degradation Biosurfactant Produced by Bacillus sp. TBM40-3)

  • 김선희;이상철;유주순;주우홍;정수열;최용락
    • Applied Biological Chemistry
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    • 제47권2호
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    • pp.170-175
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    • 2004
  • 환경친화적이며 원유 분해능이 강력한 균주를 얻기 위하여, 태백산의 토양 일부를 채취하였다. 채취한 토양 sample 로부터 crude oil을 탄소원으로 이용하는 수십 종의 균주를 분리하였으며, 분리된 균주 중 원유분해능 및 biosurfactant 생성능이 가장 우수한 균주를 선별하였다. 또한 분리된 균주의 형태학적 특성을 관찰하고 165 rDNA sequence를 통하여 Bacillus sp. TBM40-3로 동정하였다. 또한 phylogenetic tree를 이용하여 이미 동정된 다른 균주들과의 유연관계를 파악하였다. Bacillus sp. TBM40-3의 배양액의 표면장력은 최저 29 mN/m까지 감소되었고, 생산된 biosurfactant의 유화활성은 soybean oil에서 최대였으며, crude oil에서도 높은 편이였다. 유화안정성은 합성계면활성제와 비슷하거나 우수하였다. 따라서 TBM40-3에서 추출한 biosurfactant는 합성계면활성제를 대체할 수 있는 환경친화적인 생물 계면활성제로 사용될 수 있는 가능성을 보여준다.

Production of a New Biosurfactant by a New Yeast Species Isolated from Prunus mume Sieb. et Zucc.

  • Jeong-Seon Kim;Miran Lee;Dae-Won Ki;Soon-Wo Kwon;Young-Joon Ko;Jong-Shik Kim;Bong-Sik Yun;Soo-Jin Kim
    • Journal of Microbiology and Biotechnology
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    • 제33권8호
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    • pp.1023-1029
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    • 2023
  • Biosurfactants reduce surface and interfacial tension due to their amphiphilic properties and are an eco-friendly alternative for chemical surfactants. In this study, a new yeast strain JAF-11 that produces a biosurfactant was selected using drop collapse method, and the properties of the extracts were investigated. The nucleotide sequences of the strain were compared with closely related strains and identified based on the D1/D2 domain of the large subunit ribosomal DNA (LSU) and internal transcribed spacer (ITS) regions. Neodothiora populina CPC 39399T, the closest species with strain JAF-11, showed a sequence similarity of 97.75% for LSU and 94.27% for ITS, respectively. The result suggests that the strain JAF-11 represents a distinct species that cannot be assigned to any existing genus or species in the family Dothideaceae. Strain JAF-11 produced a biosurfactant reducing the surface tension of water from 72 mN/m to 34.5 mN/m on the sixth day of culture and the result of measuring the critical micelle concentration (CMC) by extracting the crude biosurfactant was found to be 24 mg/l. The molecular weight 502 of the purified biosurfactant was confirmed by measuring the fast atom bombardment mass spectrum. The chemical structure was analyzed by measuring 1H nuclear magnetic resonance (NMR), 13C NMR, and two-dimensional NMRs of the compound. The molecular formula was C26H46O9, and it was composed of one octanoyl group and two hexanoyl groups to myo-inositol moiety. The new biosurfactant is the first report of a compound produced by a new yeast strain, JAF-11.

청국장 발효 중 surfactin 생산량 품질지표 설정 (Establishment of a quality index of surfactin production during cheonggukjang fermentation)

  • 지창근;홍정화;이상현
    • 한국식품저장유통학회지
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    • 제30권2호
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    • pp.311-320
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    • 2023
  • 본 연구에서는 청국장 발효 시간별 surfactin 생산량을 신속하게 확인할 수 있는 품질지표를 선정하기 위해 물리화학적 인자들(pH, 색차, 굴절률, 흡광도, 표면장력, 단백질)을 측정하여 surfactin 생산량과 비교하였다. 일원분산분석을 통해 최대 surfactin 생산량(125.94±4.15 mg/kg)을 확인하였으며, 발효 36시간 이후부터 유의적인 증가를 나타내지 않았다. 물리화학적 인자들 중 pH, 단백질 측정은 발효 48시간까지 꾸준히 증가하는 경향을 나타내었고, 표면장력은 발효 시작 후 감소하여 24시간 이후 유의적인 변화를 나타내지 않았다. 물리적 인자인 색차, 굴절률, 흡광도는 발효 36시간에서 최대값을 나타내었으며, 36시간 이후로는 유의적인 변화가 측정되지 않았다. Surfactin 함량과 물리화학적인 인자들 간의 연계성을 분석한 결과, 색차, 굴절률, 흡광도 측정이 유의적으로 높은 유사성을 나타내었다. 따라서 청국장의 발효과정 중 surfactin 생산량을 과학적으로 규명된 지표인자를 사용함으로써 우리 전통 발효식품의 품질관리 지표로 활용함으로써 공정관리 효율성을 높일 수 있을 것으로 기대된다.