• Title/Summary/Keyword: LAC

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Microbial β-Galactosidase of Pediococcus pentosaceus ID-7: Isolation, Cloning, and Molecular Characterization

  • Lee, Ji-Yeong;Kwak, Mi-Sun;Roh, Jong-Bok;Kim, Kwang;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.598-609
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    • 2017
  • Pediococcus pentosaceus ID-7 was isolated from kimchi, a Korean fermented food, and it showed high activity for lactose hydrolysis. The ${\beta}$-galactosidase of P. pentosaceus ID-7 belongs to the GH2 group, which is composed of two distinct proteins. The heterodimeric LacLM type of ${\beta}$-galactosidase found in P. pentosaceus ID-7 consists of two genes partially overlapped, lacL and lacM encoding LacL (72.2 kDa) and LacM (35.4 kDa). In this study, Escherichia coli MM294 was used for the production of LacL, LacM, and LacLM. These three types of recombinant proteins were expressed, purified, and characterized. The specific activities of LacLM and LacL were 339 and 31 U/mg, respectively. However, activity was not detected with LacM alone. The optimal pH of LacLM and LacL was pH 7.5 and pH 7.0, and the optimal temperature of LacLM and LacL was $40^{\circ}C$ and $50^{\circ}C$, respectively. The optimal temperature changes indicate that LacLM is able to achieve higher activity at a relatively lower temperature. LacLM was strongly activated by $Mg^{2+}$, $Mn^{2+}$, and $Zn^{2+}$, which was not true for LacL. Consistent with this, EDTA strongly inactivated LacLM and LacL, but the presence of reducing agents did not dramatically alter the activity. Taken together, multiple alignment of amino acid sequences and phylogenetic analysis results of LacL and LacM of P. pentosaceus ID-7 suggest the evolution of LacL into LacLM and that the use of divalent metal ions results in higher activity.

Evaluation of Lac Cultivation in Two South-Western Districts of Bangladesh

  • Bahar, Habibullah;Islam, Tarikul;Islam, Monirul;Mannan, Abdul
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.1
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    • pp.79-82
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    • 2007
  • Lac is the dermal secretion of lac insect, Kerria lacca Kerr., which is used to make expensive natural dye, burnish, coating materials, in cosmetics and jewelry industry. Though it is very perspective industry and available of host plants over the country especially in southern districts of Bangladesh, lac cultivation is confined within some northern districts of Bangladesh. Experiment was conducted to find out the possibility of lac cultivation in two southwestern districts of Bangladesh: Khulna and Satkhira compared to Chapainowabgonj, the key lac producing district in Bangladesh, during February-October, 2006. The bark thickness, bark weight of ber plant, ratio of harvested and inoculated lac sticks, harvested raw lac, and harvested processed lac were measured and compared. Statistically, similar performance of lac cultivation was found regarding all parameters in three districts. These results recommended that lac cultivation is possible at the south-western coastal part of Bangladesh.

Characteristics and Occurance of Spontaneous Mutant in Lactobacillus casei YIT 9018 (Lactobacillus casei YIT 9018의 자연돌연변이주의 발생과 그 특성)

  • 강국희;김동운;차송일
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.463-467
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    • 1988
  • In this study characterization of spontaneous Lac$^-$ mutant of Lactobacillus casei YIT 9018 was investgated. The frequency of occuring the spontaneous Lac$^-$ mutant cells from stok culture strains was approximately 0.5% in MRS Media and 0.29% in 10% nonfat dried milk, ohterwise, that from pure Lac$^+$ colony it was 0.25% in MRS Media and 0.1% in l0% nonfat dried milk. In the physiological deference the spontaneous Lac$^-$ mutants were defective in lactose and/ or galactose-fermenting ability and also needed longer time for coagulation of nonfat dried milk than wild type.

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Influences of Cultural Temperature on Growth Rates of Lactic Acid Bacteria Isolated from Kimchi (배양온도가 김치에서 분리한 젖산균의 증식속도에 미치는 영향)

  • 소명환;이영숙
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.110-116
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    • 1997
  • Growth curves of seven strains of lactic acid bacteria isolated from Kimchi were graphed during cultivation at 1$0^{\circ}C$, 2$0^{\circ}C$, 3$0^{\circ}C$ and 4$0^{\circ}C$ in filter sterilized Chinese cabbage juice, and then lag time and generation time at each conditions were calcurated. At 3$0^{\circ}C$, the lag time of Leu. mesenteroides subsp. dextranicum was 168 min(minutes), Leu. mesenteroides subsp. mesenteroides 204 min, Leu. paramesenteroides 612 min, Lac. bavaricus 258 min, Lac. homohiochii 228 min, Lac. plantarum 270 min and Lac. brevis 264 min. And at this temperature, the generation times of Leu. mesenteroides subsp. dextranicum and Leu. mesenteroides subsp. mesenteroides were all 36 min, Lac. bavaricus 33 min, Lac. homohiochii 39 min, Lac. plantarum 66 min, Lac. brevis 42 min and Leu. paramesenteroides 162 min. As cultural temperature was lowed from 3$0^{\circ}C$ to 1$0^{\circ}C$, all strains showed remarkable prolongations in lag time and in generation time, and the prolongations were most conspicuous in Lac. plantarum. At 1$0^{\circ}C$, 2$0^{\circ}C$ and 3$0^{\circ}C$, both the lag time and the generation time of Leu. mesenteroides subsp. mesenteroides were shorter than those of Lac. plantarum. But at 4$0^{\circ}C$, this pattern was completely inverted. As a whole lower temperatures were more favorable for the growth of Leu. mesenteroides subsp. mesenteroides, while higher temperatures were for Lac. plantarum.

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An Influence of Salt Concentrations on Growth Rates of Lactic Acid Bacteria isolated from Kimchi (소금농도가 김치에서 분리한 젖산균의 증식속도에 미치는 영향)

  • 소명환;이영숙
    • The Korean Journal of Food And Nutrition
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    • v.9 no.3
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    • pp.341-347
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    • 1996
  • Growth curves of seven strains of lactic acid bacteria isolated from Kimchi were graphed, when they were cultivated at 3$0^{\circ}C$ in filter sterilized Chinese cabbage juice containing 0, 2, 4 and 6% salt, and then lag time and generation time were calculated. The shapes of growth curves were changed differently among strains, as salt concentrations were increased. The addition of 2~4% salt resulted in prolongation of lag time were most omspicuous in Leu. Paramesenteroides and Leu. Mesenteroides subsp. Dextranicum, and the next in Leu. Mesenteroides subsp. Mesenteroides,m Lac. Bavaricus and Lac. Gomohiochii, and the least in Lac. Plantarum and Lac. Brevis. And then the prolongations of generation time were most remarkable in Lac. bavaricus and Lac. Homohiochii, and the next in Leu. Mesenteroides subsp. Mesenteroides, Leu. Mesenteroides subsp. Dextranicum and Leu. Paramesenteroides, and the least in Lac. Plantarum and Lac. brevis. By increasing salt concentrations from 0 to 2%, the generation times of Leu. Cesenteroides subsp. Dextranicum and Leu. Mesenteriodes subsp. Mesenteroides were prolonged slightly, while those of Lac. Homohiochii and Lac. Brevis were not changed, and those of Lac. Plantarum, Lac. Bavaricus and Leu. Paramesenteroides were shortened slightly. As salt concentrations were increased from 2% to 4%, inversions in the order of generation time occurred among strains. As a whole, lower salt concentrations were more favorable for the growth of Leuconostoc strains, while higher salt concentrations were for Lactobacillus strains.

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Genome-Wide Identification and Characterization of Novel Laccase Genes in the White-Rot Fungus Flammulina velutipes

  • Kim, Hong-Il;Kwon, O-Chul;Kong, Won-Sik;Lee, Chang-Soo;Park, Young-Jin
    • Mycobiology
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    • v.42 no.4
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    • pp.322-330
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    • 2014
  • The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges, fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylation sites (Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1 (Asn-454), fvLac-2 (Asn-437 and Asn-455), fvLac-3 (Asn-111 and Asn-237), and fvLac4 (Asn-402 and Asn-457). In addition, the first 19~20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction analyses clearly reveal that $CuSO_4$ affects the induction and the transcription level of these laccase genes.

Lactic Acid Bacteria Isolated from Fermenting Kimchi and Their Fermentation of Chinese Cabbage Juice (김치에서 젖산균의 분리 및 이 세균들의 배추즙액 발효)

  • Shim, Sun-Taek;Kyung, Kyu-Hang;Yoo, Yang-Ja
    • Korean Journal of Food Science and Technology
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    • v.22 no.4
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    • pp.373-379
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    • 1990
  • Lactic acid bacteria(LAB) were isolated from fermenting Kimchi and were cultivated in filter-sterilized Chinese cabbage juice individually or in combination. LAB isolated were Lactobacillus leichimannii, Lac. fermentum, in addition to the already known Leuconostoc mesenteroides, Lac. plantarum, lac. brevis and pediococcus pentosaceus. Lac. leichimannii, Lac. fermentum and Lac. sake, the early lactobacilli, were high in number exceeding $10^4cells/ml$ at 0 time and multiplied up to $10^9cells/ml$ altogether at the 3rd day of kimchi fermentation. When the representative LAB were cultivated singly in Chinese cabbage juice with or without 3.0% NaCl, one strain of Leu. mesenteroides and La. leichmannii were not different in acid producing ability while the other strain of Leu. mesenteroides and Lac. fermentum Lac. plantarum, produced less acid when NaCl was present. When the bacteria in combination were cultivated in Chinese cabbage juice with 3.0% NaCl, the presence of Leu. mesenteroides was essential to eliminate the lag phase in acid production with higher amounts of acid produced than without. The total number of lactobacilli in the mixture of kimchi ingredients was about $2.9{\times}10^4 cells/ml$ while the number of Lac. plantarum was 7.3 cells/ml. The number of Lac. plantarum in individual ingredients were normally in the range between $0.0{\sim}240cells/g$ except garlic sold in ready-to-use form with $9.0{\times}10^3 cells/g$.

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발효유의 종균 L. casei YIT 9018의 자연돌연변이주의 발생 빈도 및 그 특성에 관한 연구

  • 김동운;차송일;강국희
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.513.3-514
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    • 1986
  • 유산균을 종균으로 하여 유제품을 만드는 경우에 종균의 취급관리는 매우 중요한 일이다. 종균을 취급하는 과정에서 어떤 원인으로 인하여 종균의 활력 특히 산생성력이 감퇴되어 종균으로 사용할 수 없는 경우가 발생한다. 이러한 경우의 원인을 분석하여 그 예방책을 수립하기 위한 기초 연구로써 발효유의 종균으로 사용하고 있는 L. casei YIT9018의 spontaneous lac 변이주의 발생빈도 및 이러한 Lac 변이주의 특성을 검토하였다. parent colony에서의 Lac의 발생빈도는 MRS 배지에서 400 coloney중에서 1개, 10% 환원 탈지분유에서는 984개중 1개의 배율로 나타났다. 분리된 Lac 변이주중에는 Lac Gal도 검출되었다 대부분의 Lac를 탈지유배지에 배양할 경우 10일이 지나면 우유를 응고시켰으나 Lac Gal는 30일이 지난뒤에 응고시켰다. Lac변이주를 MRS-lactose 배지의 plate에 도말하여 배양하면 4일 이후부터는 revertant가 나타났으나 Lac Gal Cell에서는 인정되지 않았다. 얻어진 Lac 변이주의 plasmid를 분리하여 parent와 비교한 결과 plasmid DNA가 소실되었음을 확인하였다.

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Identification of Potential Corynebacterium ammoniagenes Purine Gene Regulators Using the pur-lacZ Reporter in Escherichia coli

  • HAN , RI-NA;CHO, ICK-HYUN;CHUNG, SUNG-OH;HAN, JONG-KWON;LEE, JIN-HOO;KIM, SOO-KI;CHOI, KANG-YELL
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1249-1255
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    • 2004
  • This study has developed Corynebacterium ammoniagenes (c. ammoniagenes) purine gene transcriptional reporters (purF-lacZ and purE-lacZ) that function in Escherichia coli (E. coli) DH5a. After transformation of a C. ammoniagenes gDNA library into E. coli cells harboring either purF-lacZ or purE-lacZ, C. ammoniagenes clones were obtained that repress purF-lacZ and purE-lacZ gene expression. The potential purE and purF regulatory genes are homologous to the genes encoding transcription regulators, the regulatory subunit of RNA polymerase, and genes for purine nucleotide biosynthesis of various bacteria. The C. ammoniagenes purE-lacZ and purF-lacZ reporters were repressed by adenine and guanine within E. coli, indicating similarity in the regulatory mechanism of purine biosynthesis in C. ammoniagenes and E. coli. Gene regulation of pur-lacZ by adenine and guanine was partly abolished in cells expressing potential purine regulatory genes, indicating functionality of the purine gene regulators in repression of purE-lacZ and purF-lacZ. The purE-lacZ and purF-lacZ reporters can be used for the screening of genes involved in the regulation of the de novo synthesis of the purine nucleotides.

SS Lac IN OPEN CLUSTER NGC 7209 (산개성단 NGC 7209에 속한 쌍성 SS Lac)

  • 강영운
    • Journal of Astronomy and Space Sciences
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    • v.14 no.2
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    • pp.216-224
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    • 1997
  • We present IUE observations in far ultraviolet region for SS Lac in open cluster NGC 7209. The IUE short wavelength spectrum and long wavelength spectrum were merged to produce an energy distribution curve between $1200{AA}$ and $3200{AA}$. The curve has been unreddened to take out reddening effect. Then the surface temperature, gravity and abundance of SS Lac were determined based on the Kurucz model atmosphere. The photographic light curve produced before cessation of eclipse were analyzed for photometric solutions. We confirmed SS Lac is a member of open cluster NGC 7209 based on its possible distance. The main reason for the cessation of eclipse is the change of the system's orbital inclination rather than collisional merge of two stars.

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