• Title/Summary/Keyword: lactic acid bacteria(LAB)

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Evaluation and modification of alkaline lysis plasmid preparation method from Lactobacillus spp.

  • Lee, Deog-Yong;Seo, Yeon-Soo;Kang, Sang-Gyun;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.157-162
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    • 2007
  • Lactic acid bacteria (LAB) has been regarded as a useful microorganism and tried to manipulate plasmid DNA for increasing the usefulness. Although several methods have been developed to isolate plasmid DNA from Escherichia coli (E. coli), these methods were not sufficient to apply to LAB with exception of O'Sullivan's lysis method. So, we evaluated plasmid DNA extraction from LAB using general E. coli preparation methods and tried to improve the extraction yield and DNA purity by modifying O'Sullivan's alkaline lysis method. To improve the extraction yield, salt and carrier were added to precipitant and those were incubated at $-70{^{\circ}C}$. Only incubation at $-70{^{\circ}C}$ was the effective method of those modifications. Purity of plasmid DNA was improved by two times of each centrifugation and phenol/chloroform extraction. However, DNA was damaged by twice extraction with phenol/chloroform. Also, exclusion of ethidium bromide showed negative effect to purity. Additionally, it was recommended that improvement of the extraction yield may be due to centrifugation at high speed for more time and to dissolving complete DNA pellet before addition of 7.5 M ammonium acetate. Extraction using this modification produced higher quality of plasmid DNA.

Rapid and Simple Method for Isolating Plasmid DNA from Lactic acid Bacteria (유산균 Plasmid DNA의 신속 간편한 분리방법)

  • Bae, Hyung-Seok;Baek, Young-Jin;Kim, Young-Ki;Yoo, Min;Park, Moo-Young
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.289-296
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    • 1985
  • A simple procedure for rapid isolation of plasmid DNA from lactobacillus species and streptococcus species is described. Lactic acid bacteria were cultured in the TCM broth containing 0.5% glycine and plasmid DNA was isolated from cells treated with mutanolysin by alkaline-detergent lysis method. Good results for releasing and isolating plasmid DNA from lactobacillus species were obtained by treatment of cells with 30$\mu\textrm{g}$ of mutanolysin per ml at 37$^{\circ}C$ for 5 to 10 min. For the streptococcus species, the optimum conditions were slightly different. The procedure could be used for rapid characterization of plasmid DNA in Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus helveticus, Streptococcus lactis, Streptococcus faecalis, Streptococcus faecium, and Streptococcus cremoris strains. Using this procedure, plasmids isolated from $1.5m\ell$ cultures could readily be visualized in agarose gel.

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Reduction of Insulin Resistance by Momordica Charantia with Lactobacillus Acidophilus CBT-LA1 or Lactiplantibacillus Plantarum CBT-LP3 Improves Hepatosteatosis

  • Dong-Jin Kim;Ju Sung Lee;Seungwoo Kim;Sang Kyun Park;Yeo-Sang Yoon;Yougku Ryu;Myung Jun Chung
    • Microbiology and Biotechnology Letters
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    • v.52 no.3
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    • pp.275-287
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    • 2024
  • Insulin resistance is a primary risk factor for developing diabetes. However, diabetes drugs generally focus on regulating and lowering patients' blood glucose levels. In recent years, diverse materials have been evaluated to improve insulin resistance and hinder the development of diabetes. Momordica charantia extract (MCE) and lactic acid bacteria (LAB) have been considered as potential therapeutic agents against insulin resistance and hyperglycemia. In a streptozotocin-induced type 1 diabetes animal model, treatment with MCE and LAB had no effect on hyperglycemia. To evaluate the effect of MCE and LAB on insulin resistance, we chose a high-fat diet-induced insulin resistance model and co-administered MCE and Lactobacillus Acidophilus CBT-LA1, Lactiplantibacillus plantarum CBT-LP3, or Lacticaseibacillus rhamnosus CBT-LR5. MCE with CBT-LA1 or CBT-LP3 improved insulin resistance and hepatosteatosis. However, the effect of MCE and MCE with CBT-LR5 was weaker than the effect of MCE with CBT-LA1 or CBT-LP3. Momordica charantia induced insulin secretion from RIN-m5F in a dose-dependent manner. Interestingly, CBT-LA1 and CBT-LP3 enhanced the insulin secretion of MCE. These results suggest that the co-administration of MCE and a specific LAB is one approach for overcoming insulin resistance and hyperglycemia.

Characterization of Lactic Bacterial Strains Isolated from Raw Milk

  • Kim, Hyun-jue;Shin, Han-seung;Ha, Woel-kyu;Yang, Hee-jin;Lee, Soo-won
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.1
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    • pp.131-136
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    • 2006
  • During lactic acid bacteria (LAB) transit through the gastrointestinal tract, ingested microorganisms were exposed to successive stress factors, including low pH in the human stomach and in bile acid. These stress factors can be used as criteria for the selection of a viable probiotic strain. Four such strains (Lactobacillus helveticus SGU 0011, Lactobacillus pentosus SGU 0010, Streptococcus thermophilus SGU 0021 and Lactobacillus casei SGU 0020) were isolated from raw milk. When the identified LAB were exposed to synthetic gastric juice, whereas L. casei SGU 0020 and S. thermophilus SGU 0021 exhibited a 0% survival rate, L. helveticus SGU 0011 and L. pentosus SGU 0010 exhibited 60% and 95% survival rates. L. casei SGU 0020 and S. thermophilus SGU 0021 could not be examined with regard to their tolerances to artificial bile juice, as they uniformly died upon exposure. However, L. helveticus SGU 0011 and L. pentosus SGU 0010 individually survived at rates of 39% and 93%. Also, all four of these strains were confirmed to be tolerant of ten different antibiotics.

Biological and Functional Characteristics of Lactic Acid Bacteria in Different Kimchi (김치 종류에 따른 유산균의 생물학적 및 기능적 특성)

  • Ko, Kang Hee;Liu, Wenli;Lee, Hyun Hee;Yin, Jie;Kim, In Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.1
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    • pp.89-95
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    • 2013
  • Biological and functional characteristics of lactic acid bacteria (LAB) were investigated in mustard stem/leaf kimchi (MK), cabbage kimchi (CK), young radish kimchi (YRK), and cubed radish kimchi (CRK). LAB of young radish kimchi were mainly composed of bacilli in contrast to the other kimchi. 89.2% LAB isolated from all kimchi harbored plasmids. However, LAB had an average of $4.1{\pm}0.5$ plasmid bands in YRK, more than MK, CK, and CRK. Exopolysaccharides were produced by 10.9~11.1% of LAB, and were especially by LAB isolated from radish kimchi. A significant percentage of LAB (69.5%) had antibacterial activity against one sensitive strain or more. LAB from CK, YRK and CRK had antimicrobial activities against Bacillus sp., Listeria monocytogenes, and Salmonella Typhimurium, while the LAB from MK had activities against Vibrio parahaemolyticus higher than those from the other kimchi. In YRK and CRK, acid-tolerant LAB were twice as prevalent as those in MK and CK. Bile-tolerant LAB isolated from CRK were more prevalent than other kimchi. When $10^8$ CFU of LAB were added to Caco-2 cells, 12.1% of LAB isolated from all kimchi showed similar adherent activity to Lactobacillus rhamnosus GG. LAB of MK particularly adhered to Caco-2 cells, 2.0~4.1 fold higher than LAB in the other kimchi. From these results, biological and functional characteristics of LAB varied according to the type of kimchi and LAB existing in kimchi were limited to their respective species.

Isolation and Characterization of Lactobacillus buchneri Strains with High ${\gamma}$-Aminobutyric Acid Producing Capacity from Naturally Aged Cheese

  • Park, Ki-Bum;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.15 no.1
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    • pp.86-90
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    • 2006
  • Two lactic acid bacteria (LAB) with high ${\gamma}$-aminobutyric acid (GABA)-producing capacity were isolated from naturally aged cheese. Examination of the biochemical features using an API kit indicated that the two strains belonged to Lactobacillus. They were gram positive, rod-type bacteria, and fermented arabinose, melezitose, melibiose and xylose, but did not utilize cellobiose or trehalose. 16S rDNA sequencing analysis confirmed that they were Lactobacillus buchneri and Lactobacillus sp. They were accordingly named as Lactobacillus buchneri OPM-1 and Lactobacillus sp. OPM-2, and could produce GABA from MRS broth supplemented with 10 g/L of monosodium glutamate (MSG) at a productivity of 91.7 and 116.7 mg/L/hr, respectively. Cell extracts of L. buchneri OPM-1 and Lactobacillus sp. OPM-2 showed glutamate decarboxylase (GAD) activity, for which the optimum pH and temperature were 5.5 and $30^{\circ}C$, respectively.

Biological Control of Root-Knot Nematodes by Organic Acid-Producing Lactobacillus brevis WiKim0069 Isolated from Kimchi

  • Seo, Hye Jeong;Park, Ae Ran;Kim, Seulbi;Yeon, Jehyeong;Yu, Nan Hee;Ha, Sanghyun;Chang, Ji Yoon;Park, Hae Woong;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.662-673
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    • 2019
  • Root-knot nematodes (RKNs) are among the most destructive plant-parasites worldwide, and RKN control has been attempted mainly using chemical nematicides. However, these chemical nematicides have negative effects on humans and the environment, thus necessitating the search for eco-friendly alternative RKN control methods. Here, we screened nematicidal lactic acid bacteria (LAB) isolated from kimchi and evaluated their efficacy as biocontrol agents against RKNs. Of 237 bacterial strains, Lactobacillus brevis WiKim0069 showed the strongest nematicidal activity against the second-stage juveniles (J2) of Meloidogyne incognita, M. arenaria, and M. hapla and inhibited the egg hatch of M. incognita. The culture filtrate of WiKim0069 had a pH of 4.2 and contained acetic acid (11,190 ㎍/ml), lactic acid (7,790 ㎍/ml), malic acid (470 ㎍/ml), and succinic acid (660 ㎍/ml). An artificial mixture of the four organic acids produced by WiKim0069 also induced 98% M. incognita J2 mortality at a concentration of 1.25%, indicating that its nematicidal activity was derived mainly from the four organic acids. Application of WiKim0069 culture filtrate suppressed the formation of galls and egg masses on tomato roots by M. incognita in a dose-dependent manner in a pot experiment. The fermentation broth of WiKim0069 also reduced gall formation on melon under field conditions, with a higher efficacy (62.8%) than that of fosthiazate (32.8%). This study is the first report to identify the effectiveness of kimchi LAB against RKNs and to demonstrate that the organic acids produced by LAB can be used for the RKN management.

Comparison of Mcrobial and Physicochemical Properties between Pogi Kimchi and Mat Kimchi (포기김치와 맛김치의 미생물학적 및 이화학적 품질 특성 비교)

  • Moon, Eun Woo;Kim, Su-Yeon;Dang, Yun-Mi;Park, Boyeon;Park, Eun Jin;Song, Hye Yeon;Yang, Jisu;Yoon, So Ra;Seo, Hye-Young;Ha, Ji-Hyoung
    • Journal of the Korean Society of Food Culture
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    • v.34 no.2
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    • pp.217-223
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    • 2019
  • This study aimed to evaluate the characteristic differential between whole cabbage kimchi (pogi kimchi) and sliced cabbage kimchi (mat kimchi) during kimchi fermentation at $6^{\circ}C$. The difference of microbial and physicochemical properties was investigated until 6 weeks. For the changes in the microbial flora, both kimchi samples exhibited a continuous increase in total aerobic bacteria and lactic acid bacteria (LAB) population size up to 2 weeks followed by a stationary phase until 5 weeks. Interestingly, the number of LAB of mat kimchi was overall higher than that of pogi kimchi during kimchi fermentation. We speculate that mat kimchi has in a more advantageous growth condition than pogi kimchi for microbial growth because small kimchi cabbage size appropriately derives nutritional supply in order to increase the LAB growth. During lactic fermentation at $6^{\circ}C$, physicochemical changes in the pH, salinity, and titratable acidity was observed to be no significant differences between two types of kimchi. Furthermore the contents of organic acids such as oxalic acid, citric acid, malic acid, lactic acid, fumaric acid, and acetic acid was not significantly different (p>0.05) between both kimchi samples as well as the contents of total free amino acid.

Effect of Heterofermentative Lactic Acid Bacteria on the Quality of Italian Ryegrass and Whole-crop Barley Silage (Hetero 발효 유산균주 첨가가 사일리지용 이탈리안 라이그라스와 총체보리의 품질에 미치는 효과)

  • Lee, Ho Il;Choi, Yeon Jae;Mamuad, Lovelia;Kim, Eun Joong;Oh, Young Kyoon;Park, Keun Kyu;Lee, Sang Suk
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.34 no.4
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    • pp.269-276
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    • 2014
  • This study was conducted to determine the quality of italian ryegrass (IRG) and whole- crop barley (WCB) silage combined with heterofermentative lactic acid bacteria (LAB) during fermentation. Six strains of LAB (L. plantarum IMAU 70164, L. acidophilus KACC 12419, L. casei KACC 12413, L. reuteri KCTC 3594, L. buchneri KACC 12416 and L. diolivorans KACC 12385) were used in this study. L. casei and L. reuteri had the highest propionic acid production and were therefore used for fermenting the forages. The forages were fermented using monoculture and co-culture of L. casei and L. reuteri for 30, 45 and 60 days of ensiling. Addition of LAB lowered the pH of the IRG silage (p<0.05). Moisture content, lactic acid and acetic acid contents were higher (p<0.05) after addition of LAB. Water soluble carbohydrate was significantly lower (p<0.05) in WCB with a co-culture containing L. casei and L. reuteri. Propionic acid production was comparatively higher after addition of LAB to WCB on days 30, 45 and 60 while butyric acid was only detected in the IRG control on day 60. Fungi was not detected within 60 days after addition of LAB in IRG and WCB. Through this experiment, improved forage preservation was achieved using a co-culture containing L. casei and L. reuteri. WCB silage had higher propionic acid concentration and thus, it was a better forage for ensiling using co-culture of L. casei and L. reuteri.

Quality Characteristics of Fermentation Gastrodia elata Blume by Saccharifying Methods (당화방법에 따른 유산균 발효천마의 품질특성)

  • Song, Young-Eun;Choi, So-Ra;Song, Eun-Ju;Seo, Sang-Young;Lee, In-Sok;Han, Hyun-Ah;Lee, Ki-Kwon;Song, Young-Ju;Kim, Young-Hoi;Kim, Myung-Kon;Park, Shin-Young
    • The Korean Journal of Food And Nutrition
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    • v.29 no.5
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    • pp.698-705
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    • 2016
  • This study aimed to evaluate the quality characteristics of Gastrodia elata Blume fermented by lactic acid bacteria after saccharifying by 3 methods including enzyme, malt, and rice-nuruk. The lactic acid bacteria (LAB), Pediococcus inopinatus BK-3, isolated from kimchi could reduce the unpleasant taste and odor of Gastrodia elata Blume. The total acidity value of Gastrodia elata fermented by LAB on the malt and rice-nuruk extract solution for 3 days was 2.23% and 2.33%, respectively. After saccharification by malt and rice-nuruk extract solution for 3 days, the viable cell number of fermented Gastrodia elata was 9.14 log cfu/mL and 9.27 log cfu/mL, respectively. The total acidity values were increased above 3.35% by malt and rice-nuruk extract solution for 8 days. Thus, the viable cell number was the highest by malt and rice-nuruk extract solution fermentation for 3 days. The amino acid content of Gastrodia elata fermented by LAB after saccharification by malt extract solution was higher than that of other saccharifying methods. The free sugar content and p-hydroxybenzyl derivatives induced by the enzyme method were higher than those of other saccharifying methods. The overall acceptability was the highest at 4.2 point in Gastrodia elata fermented by malt extract solution.