프로바이오틱스 안전성 평가를 위한 항생제 저항성 유전자의 생물정보학적 분석법 비교

곽우리,김병용 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.1

The detection of the genome-based antibiotic resistance gene is an essential analysis process for the purpose of verifying the safety of probiotic strains, including lactic acid bacteria. In this study, 4 analysis platforms (AMRFinderPlus, staramr, rgi, ABRicate) were used for cross-comparison of 782 genomes corresponding to 19 kinds of probiotic species notified as functional foods. As a result of the analysis, the relatively fewest number of antibiotic resistance genes were detected in strains belonging to the order Lactobacillales, and antibiotic resistance genes were detected in 322 genomes used in the case of 2 types of Enterococcus genus. In addition, the presence and type of antibiotic resistance gene detection showed a lot of difference even for the same genome sequence depending on the database and analysis algorithm used by the analysis platform. These results can be confused in evaluating the potential for transmission of antibiotic resistance genes inherent in specific lactic acid bacteria and predicting potential risks that may occur in the future. Accordingly, it is judged that the antibiotic resistance gene-related analysis criteria need to be established more clearly and specifically in the safety evaluation of probiotic bacteria.

Bacillus licheniformis와 Lactobacillus casei 2단 발효에 의해 조제된 효소식품

강조은(Jo-Eun Kang),문기성(Gi-Seong Moon) 한국유산균프로바이오틱스학회 2016 Current Topic in Lactic Acid Bacteria and Probioti Vol.4 No.2

1-B-12 strain presenting α-amylase and protease activity was isolated from a local Doenjang product. The strain was identified as Bacillus licheniformis by 16S rRNA gene sequencing and named B. licheniformis 1-B-12. A combination of raw materials with unpolished rice (50%), rice bran (30%), and soybean (20%) was best for the enzyme activities. A lactic acid bacterium Lactobacillus casei GW140 was applied for co-culture with B. licheniformis 1-B-12 aiming at the production of a function-added enzyme food. Two-step fermentation, where 24 h fermentation with B. licheniformis 1-B-12 and additional 12 h fermentation with L. casei GW140 were performed, resulted in maintenance of both of the enzyme activities and high level of lactic acid bacteria cell count. Furthermore the freeze-dried enzyme food sample was stable at 25℃ for 45 d which was the last day we tested.

상황버섯(Phellinus baumii) 균사체 액체배양 포스트바이오틱스의 면역자극활성

김연숙,신현영,김훈,정은진,구자평,하은지,김가을에,정세빈,서형주,신광순,유광원 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.2

After liquid culture of Phellinus baumii (P. baumii) mycelium (LPBM) was prepared, LPBM was fractionated into A∼E fraction (A; hot-water extract of liquid culture including mycelia, B; crude polysaccharide of A, C; hot-water extract of mycelia, D; crude polysaccharide of C, and E; crude polysaccharide of culture broth) to evaluate for possibility as functional materials with immunostimulatory activity. In macrophage stimulatory activity, E fraction as postbiotics significantly increased secretion of NO and IL-12 from RAW 264.7 cells. Next, when the splenocytes of C3H/HeN mice were primary cultured, E fraction showed significantly mitogenic activity with enhancing mitogen-related cytokines (IFN-γ and TNF-α) production from splenocyte. E fraction also potently stimulated GM-CSF production from Peyer’s patch cells as well as Peyer’s patch-mediated bone marrow cell proliferation. In addition, the immunostimularoy E fraction contained neutral sugar (73.8%), uronic acid (10.6%), protein (7.8%), and polyphenol (7.5%), and mainly consisted of glucose (39.1%), galactose (21.7%), mannose (11.1%), galacturonic acid (9.9%), and arabinose (8.9%) as component sugars. In conclusion, it was demonstrated that postbiotics including exopolysaccharide fractionated from liquid culture of the P. baumii mycelium could enhanced immunostimulatory activity.

식중독균 제어를 위한 유용미생물의 항균물질 생산 및 유전체 특성 연구

김수진,신진송,박한솔,송지섭,이기원,방우석,조태진 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.1

This mini-review focused on the current advances in the development and the application of the antibacterial agents produced from probiotics strains against foodborne pathogens. Inhibitory activity of the pathogen growth could be achieved by co-culture with probiotics and/or the treatment of metabolites extracted from probiotics culture, whereas strain-dependent efficacy was mainly reported according to the bacterial species of the target pathogens. To overcome the limited antibacterial spectrum and the efficacy of the metabolites from probiotics, the recent research highlight the discovery of novel bioactive substances with broad range of the inhibitory activity of foodborne pathogens and mode-of-action which has not been reported as the major research goal. Moreover, understanding the distribution of functional and regulation genes coding the production of the antibacterial metabolites based on the bacterial genome analysis can provide the clues for the mechanisms of the pathogen control by using probiotics. Major strategies on the application of the genomics in this research area can be represented as follows: 1) functional annotation specialized for antimicrobial proteins, 2) assessment of the antibacterial effects followed by the general/functional annotation, 3) genome and metabolites analysis for the purification of antimicrobial proteins, 4) comparative genomics and the characterization of antimicrobial potential. Especially the relationship between phenotype-genotype based on genomic bioinformation with the results of the practical efficacy tests of bacterial metabolites as an active substance of disinfectants and/or preservatives can be expected to act as supportive research for broadening our knowledge regarding the key metabolic pathways associated with the production of antibacterials from probiotics.

Lactiplantibacillus plantarum의 검은콩 발효 특성 및 열처리 발효물의 항산화 효과

황운식,유청빈,이은수,이민경,박수연,박훈 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.1

This study aimed to optimize the fermentation condition of black bean using probiotic lactic acid bacteria (LAB) and to evaluate characteristics and antioxidant activity of LAB fermented and heat-treated black bean. Two LAB strains were selected by analysis of acid resistance, bile resistance, antimicrobial activity, and antioxidant activity, and identified as Lactiplantibacillus plantarum CH9 and Lactiplantibacillus plantarum SU18 by 16S rRNA sequencing. Both strains showed similar or higher acid resistance, bile resistance and antimicrobial activity, compared to Lacticaseibacillus rhamnosus GG, a commercial probiotic strain. The heat-killed cells of CH9 and SU18 strains showed significantly (p<0.05) higher DPPH and ABTS radical scavenging activities than live cells. Fermentation of black bean (30%) treated with Alcalase using the two selected strains was found to be optimal condition, increasing viable cell count of LAB up to 10.8 Log CFU/g. During the fermentation, the titratable acidity of Alcalase-treated black bean was notably increased with concomitant decrease in the pH. LAB fermentation significantly (p<0.05) increased antioxidant activity based on DPPH and ABTS radical scavenging activities as well as total phenol content. In addition, total phenol content and antioxidant activity were significantly (p<0.05) enhanced by heat processing (121C, 15 min) of the fermented products. These findings are expected to be useful for the development of various LAB-fermented foods containing heat-killed probiotics.

Identification of Predominant Lactic Acid Bacteria Associated with Kunun-Zaki and Kindirmo a Traditional Fermented Food of Nigeria

Bamgbose Timothy,Sinha Swati,Abdullahi Isa O.,Inabo Helen I.,Bello Mohammed,Kori Lokesh D.,Ametefe Elmer N.,Anvikar Anupkumar R. 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.1

Human intestinal flora is very diverse; with lactic acid bacteria (LAB) existing as part of the most vital gut microbes that improve host health. The application of LAB as a whole organism or its metabolite in the case of probiotic and bacteriocin respectively is extensive. Thus, the need to always bio-prospect for newer strains of LAB is essential. This study focused on isolating LAB from kunun-zaki and kindirmo, a fermented non-alcoholic beverage of non-dairy and dairy sources respectively, explored their physiological and biochemical properties, antibiotics sensitivity pattern and identified based on their 16S rRNA sequencing. A total of eighty isolates were selected sixty-six from kunun-zaki and fourteen from kindirmo in which 93.7% were bacilli and 6.3% were spherical in shape having 68.75% and 30% homofermentative and heterofermentative pathway respectively. All isolates have the ability to utilize glucose to produce lactic acid while their tolerance to pH 3 and salt concentration at 2%, 4% and 6.5% varied widely. Thirty-four isolates based on their physiological and biochemical properties were selected for molecular identification to ascertain their genera and species. Limosilactobacillus fermentum (68%); Lactiplantibacillus plantarum (6%) and Weissella confusa (3%) were confirmed species isolated. Thus, it was concluded that traditional fermented foods such as kunun-zaki and kindirmo are a good source to bio-prospect for LAB for product development, starter culture and probiotic study.

Aspergillus niger를 이용한 곡류발효물의 배양특성 및 α-Glucosidase 활성 저해효과

지수빈,라채훈 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.2

Liquid-state fermentation (LSF) and solid-state fermentation (SSF) were carried out to produce enzymes and β-glucan. The modified synthetic medium composition in liquid-state fermentation (LSF) was determined to be 5% (w/v) whole oat flour, 2% (w/v) tryptone, and 2% (w/v) rice bran. The different humidity conditions of oat by solid-state fermentation were evaluated in terms of maximum production of fungal biomass, amylase, protease, and β-glucan, which were 0.23 mg/g, 7,157.38 U/g, 413.67 U/g, and 8.26% (w/w), respectively, at 60% of humidity condition. Moreover, α-glucosidase inhibition activities of fermented oat with 60% humidity at 48 h were shown to be high 49.08% (w/w). Therefore, liquid- and solid-state fermentation processes were found to enhance the overall fermentation yields of oat to produce enzymes and β-glucan.

Streptococcus thermophilus KCTC14471BP의 안전성 평가 및 실크 피브로인 코팅의 면역 증진 효능

권기철,Heo Bohye,손성호,Kwon Yoo Jin,권미진,장주현,신혜정,Kim Insu,Park Sung Yurb,김경환,Kim Byoung Kook,신창훈,김병용 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.1

Probiotic strain is known to regulate the immune system by colonizing in the intestine and interact with intestinal cell receptors of lymphoid tissue. In this study, safety of Streptococcus thermophilus KCTC14471BP and silk fibroin coating effects was evaluated with respect to mucin binding abilities and immune system modulation. S. thermophilus KCTC14471BP was coated with silk fibroin by adding 1% water-soluble calcium and 0.1% silk fibroin. S. thermophilus KCTC14471BP showed the high activities of leucine arylamidase and β-galactosidase. Regarding the antibiotic resistance tests, S. thermophilus KCTC14471BP was susceptible to ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline, and chloramphenicol. S. thermophilus KCTC14471BP coated with silk fibroin showed the improved mucin binding ability from 16.1% to 71.3% and was confirmed to have no cytotoxicity against RAW 264.7 macrophage. S. thermophilus KCTC14471BP coated with silk fibroin showed dose-dependently significant increases in pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. These results suggested that S. thermophilus KCTC14471BP can be expected as a promising probiotic bacteria for applications to food industries such as fermentation or functional foods.

Leuconostoc mesenteroides CJNU 0147 균주를 적용한 발효 자몽추출물의 장내 유해세균 억제 및 비피도박테리아 증식 효과

오승준,김자이,김혜련,문기성 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.2

Grapefruit extract rich in functional substances adjusting pH 5.0 and adding sucrose (final concentration, 2%) was fermented by Leuconostoc mesenteroides CJNU 0147 strain, consequently prepared a fermented grapefruit extract containing dextran. As a result of analyzing the growth inhibitory effect on gut harmful bacterium Clostridium difficile strain using the prepared fermented grapefruit extract, the viable cell count was significantly reduced (p<0.05 vs. control for 0.5 brix as a final concentration; p<0.001 vs. control for 1.0 brix). On the other hand, the proliferative ability of the extract for Bifidobacterium breve strain, which is one of the species of the genus Bifidobacterium well known as human gut beneficial bacteria, was confirmed (p<0.001 vs. control for both 0.5 and 1.0 brix). These results indicate fermented grapefruit extract with Leu. mesenteroides CJNU 0147 strain inhibits the growth of gut harmful bacterium C. difficile and promotes the proliferation of beneficial bacterium B. breve and is expected to be used as a functional food material for gut health.

김치에서 분리한 유산균의 생리적 특성 및 지방축적억제 효과

최성원,정은아 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.2

In this study, probiotic properties such as acid resistance, bile tolerance, intestinal adhesion and fat accumulation inhibition of lactic acid bacteria isolated from kimchi were investigated in order to develop probiotics effective anti-adipogenic effects. Among the 120 types of lactic acid bacteria isolated from kimchi, 42 types of lactic acid bacteria being to acid resistance, 35 types of lactic acid bacteria being to bile resistance, and 24 types of lactic acid bacteria being to both acid resistance and bile resistance were selected. These 24 types were tested for the intestinal adhesion ability which is an essential factor of probiotics’ function and L. rhamnosus GG was used as a positive control in this test. 10 types of lactic acid bacteria having the intestinal adhesion ability were selected. The selected 10 types of lactic acid bacteria were treated in 3T3-L1 cells and intracellular fat was stained with Oil Red O reagent to measure fat accumulation. As a result, in the group treated with lactic acid bacteria OS-15, 48, 102 and 120, fat accumulation was significantly decreased compared with the non-treated group. And the strain OS-48 showed the greatest inhibitory effect on fat accumulation. The lactic acid bacterium OS-48 was identified as Lactiplantibacillus plantarum by analysis based on 16S rRNA sequencing and was named Lactiplantibacillus plantarum OS-48. Therefore, based on the results of this study, L. plantarum OS-48 is expected to be very useful as probiotics with anti-obesity effect in the food industry such as fermented food and functional food.