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한국미생물·생명공학회지 update

Microbiology and Biotechnology Letters

  • : 한국미생물생명공학회
  • : 자연과학분야  >  생물
  • : KCI등재
  • : SCOPUS
  • : 연속간행물
  • : 계간
  • : 1598-642x
  • : 2234-7305
  • : 산업미생물학회지(~2001)→한국미생물·생명공학회지(2002~)

수록정보
수록범위 : 30권1호(2002)~49권2호(2021) |수록논문 수 : 1,163
한국미생물·생명공학회지
49권2호(2021년 06월) 수록논문
최근 권호 논문
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KCI등재 SCOPUS

1Isolation of Lactic Acid Bacteria with Anti-MRSA Bacteriocin Activity and Characterization of the Bacteriocin Product

저자 : Byeong-ki Ahn , Kyung-cheol Min , Sang-hyun Cho , Dong-geun Lee , Andre Kim , Sang-hyeon Lee

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 131-137 (7 pages)

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This study aimed at isolating lactic acid bacteria with anti-MRSA (methicillin-resistant Staphylococcus aureus) bacteriocin activity from fermented shrimp. We selected three strains, named Weissella sp. S1, S2, and S3, using analysis based on 16S rRNA gene sequences. All strains showed appropriate growth in an MRS medium containing 5% (w/v) NaCl and showed antibacterial activities against Bacillus cereus, Escherichia coli, Staphylococcus aureus, and MRSA. The strains exhibited similar growth rates at 0-5% NaCl, with approximate reduction in growth rate observed at 9% NaCl. Weissella sp. S1, S2, and S3 exhibited maximum growth rates at pH 7, 9, and 8, respectively. The crude bacteriocin was prepared from Weissella sp. S3 and subjected to characterization. The remaining activities after 30 min of exposure at each temperature were 100%, beyond 75%, and 49% at 4℃ and 37℃, 50℃ and 70℃, and 100℃, respectively. The remaining activities after 24 h of exposure at each pH were 100%, 75%, and 49% at pH 3 and 5, 7 and 9, and 10, respectively. Use of 50% (v/v) ethanol or isopropanol treatment did not diminish the antibacterial activity of the bacteriocin, while the 50% (v/v) hexane treatment reduced the activity by 51%. The molecular weight of the bacteriocin was nearly 6 kDa that was quantified using tricine-SDS-PAGE. Our findings suggest that Weissella sp. S3 may be considered a probiotic and useful source of antimicrobial substances in the development of bio-preservatives for food or in MRSA treatment.

KCI등재 SCOPUS

2Apigenin Ameliorates Oxidative Stress-induced Neuronal Apoptosis in SH-SY5Y Cells

저자 : Yeo Jin Kim , Eun Ju Cho , Ah Young Lee , Weon Taek Seo

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 138-147 (10 pages)

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The overproduction of reactive nitrogen species (RNS) and reactive oxygen species (ROS) causes oxidative damage to neuronal cells, leading to the progression of neurodegenerative diseases. In this study, we determined the nitric oxide radical (NO), hydroxyl radical (·OH), and superoxide anion radical (O2 -) scavenging activities of apigenin. Our results showed that apigenin exhibited remarkable, concentration-dependent ·OH, O2 -, and NO radical scavenging activities. Particularly, apigenin indicated the strongest ·OH radical scavenging activity with 93.38% in the concentration of 100 μM. Furthermore, we also investigated the protective effects of apigenin against hydrogen peroxide (H2O2)-induced oxidative stress in SH-SY5Y cells. The H2O2 treatment resulted in a significant decrease in cell viability, as well as an increase in lactate dehydrogenase (LDH) release and ROS production compared with the H2O2-nontreated SH-SY5Y cells. However, the cell viability significantly increased in the apigenin-treated group, as well as inhibited ROS generation and LDH release compared with the H2O2-induced control group. To elucidate the protective mechanisms of apigenin against oxidative stress in SH-SY5Y, we analyzed the apoptosis-related protein expression. The apigenin treatment resulted in the downregulated expression of apoptosis-related protein markers, such as cytochrome C, cleaved caspase-3, poly (ADP)-ribose polymerase (PARP), and B-cell lymphoma 2-associated X (Bax), as well as the upregulated expression of anti-apoptosis markers such as B-cell lymphoma 2 (Bcl-2). In this study, we report that apigenin exhibits a neuroprotective effect against oxidative stress in SH-SY5Y cells. These results suggest that apigenin may be considered as a potential agent for neurodegenerative disease prevention.

KCI등재 SCOPUS

3The Effect of Growth Condition on a Soluble Expression of Anti-EGFRvIII Single-chain Antibody in Escherichia coli NiCo21(DE3)

저자 : Kartika Sari Dewi , Ratna Annisa Utami , Hariyatun Hariyatun , Riyona Desvy Pratiwi , Dian Fitria Agustiyanti , Asrul Muhamad Fuad

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 148-156 (9 pages)

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Single-chain antibodies against epidermal growth factor receptor variant III (EGFRvIII) are potentially promising agents for developing antibody-based cancer treatment strategies. We described in our previous study the successful expression of an anti-EGFRvIII scFv antibody in Escherichia coli. However, we could also observe the formation of insoluble aggregates in the periplasmic space, limiting the production yield of the active product. In the present study, we investigated the mechanisms by which growth conditions could affect the expression of the soluble anti-EGFRvIII scFv antibody in small-scale E. coli NiCo21(DE3) cultures, attempting to maximize production. The secreted scFv molecules were purified using Ni-NTA magnetic beads and protein characterization was performed using SDS-PAGE and western blot analyses. We used the ImageJ software for protein quantification and determined the antigen-binding activity of the scFv antibody against the EGFRvIII protein. Our results showed that the highest percentage of soluble scFv expression could be achieved under culture conditions that combined low IPTG concentration (0.1 mM), low growth temperature (18℃), and large culture dish surface area. We found moderate-yield soluble scFv production in the culture medium after lactose-mediated induction, which was also beneficial for downstream protein processing. These findings were confirmed by conducting western blot analysis, indicating that the soluble, approximately 30-kDa scFv molecule was localized in the periplasm and the extracellular space. Moreover, the antigen-binding assay confirmed the scFv affinity against the EGFRvIII antigen. In conclusion, our study reveals that low-speed protein expression is preferable to obtain more soluble anti-EGFRvIII scFv protein in an E. coli expression system.

KCI등재 SCOPUS

4Improving the Viability of Freeze-dried Probiotics Using a Lysine-based Rehydration Mixture

저자 : Karina Arellano , Haryung Park , Bobae Kim , Subin Yeo , Hyunjoo Jo , Jin-hak Kim , Yosep Ji , Wilhelm H. Holzapfel

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 157-166 (10 pages)

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The probiotic market is constantly continuing to grow, concomitantly with a widening in the range and diversity of probiotic products. Probiotics are defined as live microorganisms that provide a benefit to the host when consumed at a proper dose; the viability of a probiotic is therefore of crucial importance for its efficacy. Many products undergo lyophilization for maintaining their shelf-life. Unfortunately, this procedure may damage the integrity of the cells due to stress conditions during both the freezing and (vacuum-) drying process, thereby impacting their functionality. We propose a lysine-based mixture for rehydration of freeze-dried probiotics for improving their viability during in vitro simulated gastric and duodenum stress conditions. Measurement of the zeta potential served as an indicator of cell integrity and efficacy of this mixture, while functionality was estimated by adhesion to a human enterocyte-like Caco-2 cell-line. The freeze-dried bacteria exhibited a significantly different zeta potential compared to fresh cultures; however, this condition could be restored by rehydration with the lysine mixture. Recovery of the surface charge was found to influence adhesion ability to the Caco-2 cell-line. The optimum lysine concentration of the formulation, designated “Zeta-bio”, was found to be 0.03 M for improving the viability of Lactiplantibacillus plantarum Lp-115 by up to 13.86% and a 7-strain mixture (400B) to 41.99% compared to the control rehydrated with distilled water. In addition, the lysine Zeta-bio formulation notably increased the adherence ability of lyophilized Lp-115 to the Caco-2 cell-line after subjected to the in vitro stress conditions of the simulated gastrointestinal tract passage.

KCI등재 SCOPUS

5A Study on the Tyrosinase Inhibitory and Antioxidant Effect of Microalgae Extracts

저자 : Keunho Ji , Yeeun Kim , Young Tae Kim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 167-173 (7 pages)

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Reactive oxygen species (ROS) disrupt the cellular redox balance, exert cytotoxic effects, and consequently promote the development of various diseases in humans. Previous studies have reported that antioxidants counteract the adverse effects of ROS. Several studies examine the whitening effects of various agents based on their ability to inhibit tyrosinase activity. Tyrosinase is a critical enzyme involved in the synthesis of melanin, which protects the skin against radiation. Various agents exhibiting antioxidant and tyrosinase inhibitory activities have been synthesized. However, these synthetic drugs are associated with toxicity, decreased safety, and poor skin penetration in vivo, which has limited the clinical application of synthetic drugs. This study examined the antioxidant and tyrosinase inhibitory activities of some microalgae. The methanol, dichloromethane, and ethyl acetate extracts of four microalgal species (Tetraselmis tetrathele, Dunaliella tertiolecta, Platymonas sp., and Chaetoceros simplex) were prepared. The physiological and whitening effects of microalgal extracts were investigated by measuring the antioxidant and tyrosinase inhibitory activities. The ethyl acetate extract of D. tertiolecta exhibited the highest antioxidant and tyrosinase inhibitory activities. Future studies must focus on examining the whitening effects of microalgae on cell lines to facilitate the development of microalga-based therapeutics for skin diseases, functional health foods, and whitening agents. Thus, microalgae have potential applications in the pharmaceutical, food, and cosmetic industries.

KCI등재 SCOPUS

6Biosynthesis of Three Chalcone β-D-glucosides by Glycosyltransferase from Bacillus subtilis ATCC 6633

저자 : Yinuo Fei , Yan Shao , Weiwei Wang , Yatian Cheng , Boyang Yu , Xiaorong He , Jian Zhang

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 174-180 (7 pages)

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Chalcones exhibit multiple biological activities. Various studies have attempted to modify the structure of chalcones with a special focus on the addition of substituents to the benzene rings. However, these chemical modifications did not improve the water solubility and bioavailability of chalcones. Glycosylation can markedly affect the physical and chemical properties of hydrophobic compounds. Here, we evaluated the ability of a highly promiscuous glycosyltransferase (GT) BsGT1 from Bacillus subtilis ATCC 6633 to biosynthesize chalcone glucosides. Purified BsGT1 catalyzed the conversion of 4'-hydroxychalcone (compound 1), 4'-hydroxy-4-methylchalcone (compound 2), and 4-hydroxy-4'-methoxychalcone (compound 3), into chalcone 4'-O-β-D-glucoside (compound 1a), 4-methylchalcone 4'-O-β-D-glucoside (compound 2a), and 4'- methoxychalcone 4-O-β-D-glucoside (compound 3a), respectively. To avoid the addition of expensive uridine diphosphate glucose (UDP-Glc), a whole-cell biotransformation system was employed to provide a natural intracellular environment for in situ co-factor regeneration. The yields of compounds 1a, 2a, and 3a were as high as 90.38%, 100% and 74.79%, respectively. The successful co-expression of BsGT1 with phosphoglucomutase (PGM) and UDP-Glc pyrophosphorylase (GalU), which are involved in the biosynthetic pathway of UDP-Glc, further improved the conversion rates of chalcones (the yields of compounds 1a and 3a increased by approximately 10%). In conclusion, we demonstrated an effective whole-cell biocatalytic system for the enzymatic biosynthesis of chalcone β-D-glucoside derivatives.

KCI등재 SCOPUS

7Optimization, Purification, and Characterization of Haloalkaline Serine Protease from a Haloalkaliphilic Archaeon Natrialba hulunbeirensis Strain WNHS14

저자 : Rania S Ahmed , Amira M Embaby , Mostafa Hassan , Nadia A Soliman , Yasser R Abdel-fattah

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 181-191 (11 pages)

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The present study addresses isolation, optimization, partial purification, and characterization of a haloalkaline serine protease from a newly isolated haloarchaeal strain isolated from Wadi El Natrun in Egypt. We expected that a two-step sequential statistical approach (one variable at a time, followed by response surface methodology) might maximize the production of the haloalkaline serine protease. The enzyme was partially purified using Hiprep 16/60 sephacryl S-100 HR gel filtration column. Molecular identification revealed the newly isolated haloarchaeon to be Natrialba hulunbeirensis strain WNHS14. Among several tested physicochemical determinants, casamino acids, KCl, and NaCl showed the most significant effects on enzyme production as determined from results of the One-Variable-At-A-time (OVAT) study. The Box- Behnken design localized the optimal levels of the three key determinants; casamino acids, KCl, and NaCl to be 0.5% (w/v), 0.02% (w/v), and 15% (w/v), respectively, obtaining 62.9 U/ml as the maximal amount of protease produced after treatment at 40℃, and pH 9 for 9 days with 6-fold enhancement in yield. The enzyme was partially purified after size exclusion chromatography with specific activity, purification fold, and yield of 1282.63 U/mg, 8.9, and 23%, respectively. The enzyme showed its maximal activity at pH, temperature, and NaCl concentration optima of 10, 75℃, and 2 M, respectively. Phenylmethylsulfonyl fluoride (PMSF, 5 mM) completely inhibited enzyme activity.

KCI등재 SCOPUS

8Role of Organic Spices in the Preservation of Traditionally Fermented Kunun-zaki

저자 : Williana , N. Mokoshe , Babasola , A. Osopale , Cajethan , O. Ezeamagu , Stephen O. Fapohunda

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 192-200 (9 pages)

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Kunun-zaki, produced by submerged fermentation of a combination of millet and sorghum, is a popular beverage in Northern Nigeria. Owing to the nature of the process involved in its production, kunun-zaki is highly susceptible to contamination by food spoilage microorganisms, leading to inconsistent quality and short shelf-life. In this study, we investigated various food spices, including cinnamon, garlic, and nutmeg, as potential preservatives that could be used to extend kunun-zaki shelf-life. Kunun-zaki varieties were fermented with each of these spices mentioned above and subjected to bacterial, nutritional, sensory, and quality maintenance assessments (using a twelve-member sensory panel to evaluate the organoleptic properties of kunun-zaki). Bacterial counts in the final products ranged between 105-7 CFU/ml. We identified two bacterial genera, Weissella and Enterococcus, based on partial 16S rRNA gene amplicon sequencing. Three amino acids, namely leucine, aspartate, and glutamate, were abundant in all kunun-zaki varieties, while the total essential amino acid content was above 39%, suggesting that kunun-zaki could potentially be considered as a protein-rich food source both for infants and adults. The kunun-zaki products were also rich in carbohydrates, crude proteins, ash, crude fiber, and fat, with contents estimated as 81-84, 8-11, 0.8-4.0, 2.9- 3.58, and 5.1-6.3%, respectively. However, this nutritional content depreciated rapidly after 24 h of storage, except for kunun-zaki fermented with garlic, which its crude protein and fat content was maintained for up to 48 h. Our results revealed that organic spices increased the nutritional content of the kunun-zaki varieties and could be potentially be used as natural preservatives for enhancing the kunun-zaki shelf-life. However, garlic might be considered a better alternative based on our preliminary investigation. The presence of the isolated microorganisms in the analyzed kunun-zaki samples should be highlighted to raise awareness on the possible health hazards that could arise from poor handling and processing techniques.

KCI등재 SCOPUS

9Effect of Stevia rebaudiana on the Bioactive Compounds from Agarwood Leaf (Aquilaria spp.) by Lactic Fermentation and Spray Drying

저자 : Lieu My Dong , Doan Trung Nam , Tran Thi Phuong , Dang Kim Thuy

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 201-209 (9 pages)

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Agarwood (Aquilaria spp) has high economic value. However, essential oil production from agarwood is a time-consuming process. Additionally, agarwood leaves have not been utilized even though they contain various bioactive ingredients. In this study, agarwood leaves were fermented using Lactobacillus plantarum ATCC 8014 with or without Stevia (4, 8, and 12%; v/v). The fermented fluid was mixed with maltodextrin (15%; w/v) and subjected to spray drying (inlet temperature, 120℃; outlet temperature, 65-70℃). The contents of polyphenols, polysaccharides, saponins, and flavonoids and the viability of L. plantarum were determined. Fermentation enhanced the levels of bioactive compounds. The contents of polyphenol (69.19 ± 4.05 mg GAE/g of sample), polysaccharide (20.75 ± 0.98 mg GE/g of sample), saponin (305.23 ± 4.21 mg OAE/g of sample), and flavonoid (7.86 ± 0.72 mg QE/g of sample), and the viability of L. plantarum (8.72 ± 0.17 log CFU/ml) were markedly upregulated in the samples containing Stevia (12%; v/v). This indicated that the supplementation of Stevia during fermentation decreases the fermentation time (9 h), upregulates bioactive compound production in agarwood leaves, enhances microencapsulation during spray drying, and increases the viability of L. plantarum under simulated gastric digestion conditions.

KCI등재 SCOPUS

10CRISPR/CAS9을 이용하여 lipid elongation gene의 과발현을 통한 효모의 에탄올 발효능 개선

저자 : 김진아 , 정귀택

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 2호 발행 연도 : 2021 페이지 : pp. 210-216 (7 pages)

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본 연구에서는 CRISPR/CAS9을 이용하여 S. cerevisiae의 ACC1, ELO1, OLE1 유전자의 프로모터를 TEF1으로 교체하여 그 발현량을 증가시키고 그에 따른 에탄올에 대한 저항성과 생산량 변화를 확인하였다. 18% 에탄올이 함유된 YPD 배지에서 control을 제외하고 유전자 과발현을 일으킨 mutant 균주 모두가 24시간까지 viable하게 생존하는 것을 확인하였다. 에탄올 발효에서는 유전자 과발현 균주 모두가 에탄올 수율에서 ACC1 과발현 균주가 428.18 ± 0.29 mg/g, ELO1 과발현 균주는 416.15 ± 4.3 mg/g, OLE1 과발현 균주는 430.55 ± 6.00 mg/g에 도달하였으며, 이는 control의 수율인 400.26 ± 0.42 mg/g 보다 높은 수준에 도달하였다. 이 결과는 높은 농도의 에탄올에서 탄소 사슬이 긴 불포화지방산의 비율이 증가한다는 연구결과가 역 또한 성립한다는 것을 증명하였다. ELO1의 과발현은 elongation of fatty acid protein의 생산 증가를 불러 일으킨다. 또한 OLE1도 acyl-CoA desaturase 효소의 활성을 증대시킨다. TEF1이라는 strong promoter를 이용한 이번 실험에서 ELO1 과발현 균주가 OLE1 과발현 균주보다 S. cerevisiae의 에탄올 저해 감소와 발효에 긍정적인 영향을 미침을 확인하였다.


This study aimed to enhance ethanol productivity of Saccharomyces cerevisiae through genome editing using CRISPR/CAS9. To increase ethanol productivity, ACC1, ELO1, and OLE1 were overexpressed in S. cerevisiae using the CRISPR/CAS9 system. The strains overexpressing ACC1, ELO1, and OLE1 survived up to 24 h in YPD medium supplemented with 18% ethanol. Moreover, the ethanol yields in strains overexpressing ACC1 (428.18 mg ethanol/g glucose), ELO1 (416.15 mg ethanol/g glucose), and OLE1 (430.55 mg ethanol/g glucose) were higher than those in the control strains (400.26 mg ethanol/g glucose). In conclusion, the overexpression of these genes increased the viability of S. cerevisiae at high ethanol concentrations and the ethanol productivity without suppressing glucose consumption.

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