• Title/Summary/Keyword: proteolytic bacteria

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Characterization of the Biodiversity of the Spoilage Microbiota in Chicken Meat Using Next Generation Sequencing and Culture Dependent Approach

  • Lee, Hee Soo;Kwon, Mirae;Heo, Sunhak;Kim, Min Gon;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.37 no.4
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    • pp.535-541
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    • 2017
  • This study investigated the psychrotrophic bacteria isolated from chicken meat to characterize their microbial composition during refrigerated storage. The bacterial community was identified by the Illumina MiSeq method based on bacterial DNA extracted from spoiled chicken meat. Molecular identification of the isolated psychrotrophic bacteria was carried out using 16S rDNA sequencing and their putrefactive potential was investigated by the growth at low temperature as well as their proteolytic activities in chicken meat. From the Illumina sequencing, a total of 187,671 reads were obtained from 12 chicken samples. Regardless of the type of chicken meat (i.e., whole meat and chicken breast) and storage temperatures ($4^{\circ}C$ and $10^{\circ}C$), Pseudomonas weihenstephanensis and Pseudomonas congelans were the most prominent bacterial species. Serratia spp. and Acinetobacter spp. were prominent in chicken breast and whole chicken meat, respectively. The 118 isolated strains of psychrotrophic bacteria comprised Pseudomonas spp. (58.48%), Serratia spp. (10.17%), and Morganella spp. (6.78%). All isolates grew well at $10^{\circ}C$ and they induced different proteolytic activities depending on the species and strains. Parallel analysis of the next generation sequencing and culture dependent approach provides in-depth information on the biodiversity of the spoilage microbiota in chicken meat. Further study is needed to develop better preservation methods against these spoilage bacteria.

Proteolytic System of Streptococcus thermophilus

  • Rodriguez-Serrano, G.M.;Garcia-Garibay, M.;Cruz-Guerrero, A.E.;Gomez-Ruiz, L.;Ayala-Nino, A.;Castaneda-Ovando, A.;Gonzalez-Olivares, L.G.
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1581-1588
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    • 2018
  • The growth of lactic acid bacteria (LAB) generates a high number of metabolites related to aromas and flavors in fermented dairy foods. These microbial proteases are involved in protein hydrolysis that produces necessary peptides for their growth and releases different molecules of interest, like bioactive peptides, during their activity. Each genus in particular has its own proteolytic system to hydrolyze the necessary proteins to meet its requirements. This review aims to highlight the differences between the proteolytic systems of Streptococcus thermophilus and other lactic acid bacteria (Lactococcus and Lactobacillus) since they are microorganisms that are frequently used in combination with other LAB in the elaboration of fermented dairy products. Based on genetic studies and in vitro and in vivo tests, the proteolytic system of Streptococcus thermophilus has been divided into three parts: 1) a serine proteinase linked to the cellular wall that is activated in the absence of glutamine and methionine; 2) the transport of peptides and oligopeptides, which are integrated in both the Dpp system and the Ami system, respectively; according to this, it is worth mentioning that the Ami system is able to transport peptides with up to 23 amino acids while the Opp system of Lactococcus or Lactobacillus transports chains with less than 13 amino acids; and finally, 3) peptide hydrolysis by intracellular peptidases, including a group of three exclusive of S. thermophilus capable of releasing either aromatic amino acids or peptides with aromatic amino acids.

Screening of Antimicrobial Activity and Proteolytic Enzyme Stability of Extract of the Blue Mussel Mytilus edulis (진주담치(Mytilus edulis) 추출물의 항균활성 및 단백질 분해효소에 대한 안정성 탐색)

  • Lee, Ji-Eun;Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.3
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    • pp.280-286
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    • 2021
  • This study was performed to screen the antimicrobial activities and proteolytic enzyme stability of the acidified extract of the Blue mussel Mytilus edulis. The acidified extract showed potent antimicrobial activities against Gram-positive bacteria, Bacillus subtilis, and Gram-negative bacteria, Escherichia coli D31, but had no activity against Candida albicans. Treatment of extract with trypsin completely abolished all or significant antibacterial activity against the tested bacteria, but slightly decreased antimicrobial activity against B. subtilis, and treatment of extract with chymotrypsin retained almost antibacterial activity against the tested bacteria except for E. coli D31. To confirm the additional enzyme stability of the extract, antimicrobial activity of the extract was tested after treated with several enzymes. Enzymes treated extract showed potent antimicrobial activity against B. subtilis and its activity was also retained for 5 h after trypsin treatments. Non-proteinaceous materials in the acidified extract also showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. All our results indicate that mussel extract might contain the proteinaceous or non-proteinaceous antibacterial materials target not bacterial membrane but intracellular components. These results could be used to develop mussel extract as an additive for the improvement of stability or antimicrobial activity of antibiotics against specific bacteria.

A Study on Growth Condition and Proteolytic Enzyme of Halobacterium halobium (Halobacterium halobium 의 생육조건 및 Protease 에 관한 연구)

  • 민윤식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.5
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    • pp.856-862
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    • 1994
  • In salt-preserved foods of every kinds, it was examined the growth condition of halophilic bacteria that induced a change of colour, taste, nutritive substance, a production condition of enzyme and a character of crude enzyme. Used bacteria is H. halobium a kind of extremely halophilic bacteria, and the required of optimum culture needed a quite long time of crude enzyme production is 168 hours. Optimum pH is about 7-7.5, so the traditional food of such neutrality pH as soybean paste and soy sauce particularly come into trouble because the growth can flourish in neutrality or alkaliescence, and the crude enzyme also appeared that best activation between pH 6 and pH 8. The optimum temperature is about 37$^{\circ}C$, the optimum temperature of enzyme is about 40 $^{\circ}C$ and the temperature stability is settled for 15 minutes and it is completely inactivated at 10 minutes. In the influence of each metal ion, Fe++ and Mn++ a stimulated the growth of H.halobium and the activation of enzyme, Cu++ and Zn++ were identified that made the growth and the activation of enzyme inhibit.

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Influences of Proteolytic Ability of Lactic Acid Bacteria on Acid Production and Precipitates Occurrence in Liquid Yogurt Preparation (액상발효유 제조시 유산균 Starter의 단백질 분해능이 산생성 및 침전발생에 미치는 영향)

  • 소명환
    • Microbiology and Biotechnology Letters
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    • v.12 no.4
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    • pp.285-291
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    • 1984
  • In making liquid yoghurt, the influences of proteolytic ability of lactic acid bacteria on acid production and on protein stability were investigated. L. bulgaricus CH-2, L. helviticus IAM 1042 and L. jugurti 3048 showed a comparatively high proteolytic activity in milk, while L. casei YIT 9018 did not show any marked proteolysis. Starter organisms having high proteolytic ability showed more rapid growth and acid production than those having low ability in milk. The most active proteolysis occurred during logarithmic growth phase of yogurt organisms, and most of the proteolysis took place in the first 24-48 hrs of incubation. Highly proteolysed yogurts made by L. bulgaricus CH-2, L. jugurti 3048, L. helviticus IAM 1042, L. acidophilus L-54 and L. casei 3012 had low protein solubility at pH 3.5 and had much protein precipitates during storage of product, but those having little protein hydrolysates made by L. casei YIT 9018 or artificial acidification showed no precipitation during keeping.

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Studies on the Microflora and Enzyme Activity in Processing of Accelerated Low Salt-Fermented Anchovy by adding koji (코오지를 이용한 속성 저식염 멸치젓의 미생물상과 효소활성)

  • 백승화;임미선;김동한
    • The Korean Journal of Food And Nutrition
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    • v.9 no.4
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    • pp.392-397
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    • 1996
  • To produce low salt fermented anchovy by an accelerated method with Asp. oryzae and Bacillus sp. koji, enzyme activity and variation of microflora during the 60 day fermentation were examined. Bacterial counts changed a little during the fermentation with the highest on day 40 for proteolytic and anaerobic bacteria and on day 20 for aerobic bacteria. Proteolytic, lipolytic, aerobic, and anaerobic bacteria counts were higher in the Bacillus sp. koji added anchovy paste than in others. The protease and lipase activities reached the highest point on day 20 and 30, respectively, and decreased gradually afterwards. The protease activity was higher in Asp. oryzae koji than in bacillus sp. koji, but the lipase activity was to the contrary.

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Characterization of the Strong Proteolytic Bacteria Isolated from Low Salt Fermented Anchovy and of Protease Produced by that Strain (저식염멸치젓에서 분리한 단백질분해력이 강한 세균 및 생산된 단백분해효소의 특성)

  • CHA Yong-Jun;LEE Eung-Ho;LEE Kang-Hee;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.2
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    • pp.71-79
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    • 1988
  • For the purpose of producing low salt fermented anchovy by accelerated method with a strong proteolytic bacteria, in this study, a strong proteolytic bacterium was isolated from low salt fermented anchovy and its bacteriological characteristics and properties of protease were experimented. The results obtained were as fellows : three proteolytic bacteria, Aeromonas anaerogenes Barillus subtilis and Staphylococcus saprophyticus were isolated from low salt fermented anchovy($4\%\;of\;salt,\;4\%\;of\;KCl,\;0.5\%\;of\;lactic\;acid,\;6\%$of sorbitol and $4\%$ of alcohol extract of red pepper) after 40 days fermentation. Among these strains, which grow best at $30^{\circ}C$, pH 7.0, B. subtilis was found the best proteolytic strain and benefit for industrial use as shown $0.95\;hr^{-1}$ of specific growth rate, $89{\mu}g-Tyr/hr.ml$ of maximum activity after 12 hrs culture in TPY broth. The protease produced by by B. subtilis showed maximum activity at $35^{\circ}C$, pH 7.0, and molecular weight was estimated to be 23,000 by Sephadex G-100 filtration, and it was supposed to be a kind of metal chelator sensitive neutral protease from the results of strong sensitivity against EDTA, o-phenanthroline and metal ions such as $Cu^{2+},\;Ni^{2+},\;Fe^{2+}.Km$ value of that by method of Lineweaver-Burk was determinded to be $0.73\%$ for casein as a substrate.

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Recent Advances in Biotechnology of Rumen Bacteria - Review -

  • Forsberg, C.W.;Egbosimba, E.E.;MacLellan, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.93-103
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    • 1999
  • Recent advances in the biotechnology of ruminal bacteria have been made in the characterization of enzymes involved in plant cell wall digestion, the exploration of mechanisms of gene transfer in ruminal bacteria, and the development of vectors. These studies have culminated in the introduction and expression of heterologous glucanase and xylanase genes and a fluoroacetate dehalogenase gene in ruminal bacteria. These recent studies show the strategy of gene and vector construction necessary for the production of genetically engineered bacteria for introduction into ruminants. Molecular research on proteolytic turnover of protein in the rumen is in its infancy, but a novel protein high in essential amino acids designed for intracellular expression in ruminal organisms provides an interesting approach for improving the amino acid profile of ruminal organisms.

Isolation of Bacteria from Jeotgal Using High-salt-content Media and Their Growths in High-salt Condition (고염에서 생장하는 젓갈 유래 Bacteria의 분리 및 고염에서의 생육 특성)

  • An, Doo-Hyun;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.294-300
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    • 2011
  • Proteolytic bacteria were isolated from Myeolchi-jeotgal and Saeu-jeotgal using high-salt-content media and their growths in the media containing 25% NaCl were monitored to draw the role of bacteria in the ripening of jeotgal. The most populous genus in Myeolchi-jeotgal detected on agar media with 15% NaCl was Bacillus and its relatives, while the most populous in Saeu-jeotgal was Staphylococcus. Among the isolates, Virgibacillus halodenitrificans from Myeolchi-jeotgal and Halobacillus trueperi from Saeu-jeotgal showed proteinase activities. The species from Myeolchi-jeotgal showed proteinase activity on the agar media with 8% NaCl were similar to those isolated from the media with 15% NaCl. The dominant of Myeolchi-jeotgal isolated at the 15% NaCl concentration may be involved in the proteolysis. The proteolytic species from Saeu-jeotgal on the agar media with 8% NaCl were the genera Bacillus, Salinicoccus, and Salimicrobium those were not the dominants at 15% NaCl condition. The dominant isolates from Saeu-jeotgal on agar media with 15% NaCl may not be involved in the proteolysis of Saeu-jeotgal. Vb. halodenitrificans and Staphylococcus equorum, the dominant species from Myeolchi-jeotgal and Saeu-jeotgal, showed growths at the nutrient broth containing 25% NaCl. They may play a significant role in the ripening of jeotgal and have a high possibility to be used as the starter.

Angiotensin-I-Converting Enzyme Inhibitory Peptides in Goat Milk Fermented by Lactic Acid Bacteria Isolated from Fermented Food and Breast Milk

  • Rubak, Yuliana Tandi;Nuraida, Lilis;Iswantini, Dyah;Prangdimurti, Endang
    • Food Science of Animal Resources
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    • v.42 no.1
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    • pp.46-60
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    • 2022
  • In this study, angiotensin-I-converting enzyme inhibitory (ACEI) activity was evaluated in fermented goat milk fermented by lactic acid bacteria (LAB) from fermented foods and breast milk. Furthermore, the potential for ACEI peptides was identified in fermented goat milk with the highest ACEI activity. The proteolytic specificity of LAB was also evaluated. The 2% isolate was inoculated into reconstituted goat milk (11%, w/v), then incubated at 37℃ until pH 4.6 was reached. The supernatant produced by centrifugation was analyzed for ACEI activity and total peptide. Viable cell counts of LAB and titratable acidity were also evaluated after fermentation. Peptide identification was carried out using nano liquid chromatography mass spectrometry (LC-MS/MS), and potential as an ACEI peptide was carried out based on a literature review. The result revealed that ACEI activity was produced in all samples (20.44%-60.33%). Fermented goat milk of Lc. lactis ssp. lactis BD17 produced the highest ACEI activity (60.33%; IC50 0.297±0.10 mg/mL) after 48 h incubation, viable cell counts >8 Log CFU/mL, and peptide content of 4.037±0.27/mL. A total of 261 peptides were released, predominantly derived from casein (93%). The proteolytic specificity of Lc. lactis ssp. lactis BD17 through cleavage on the amino acid tyrosine, leucine, glutamic acid, and proline. A total of 21 peptides were identified as ACEI peptides. This study showed that one of the isolates from fermented food, namely Lc. lactis ssp. lactis BD17, has the potential as a starter culture for the production of fermented goat milk which has functional properties as a source of antihypertensive peptides.