• Title/Summary/Keyword: Selective hydrolysis

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Construction and Analysis of Food-Grade Lactobacillus kefiranofaciens β-Galactosidase Overexpression System

  • He, Xi;Luan, MingJian;Han, Ning;Wang, Ting;Zhao, Xiangzhong;Yao, Yanyan
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.550-558
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    • 2021
  • Lactobacillus kefiranofaciens contains two types of β-galactosidase, LacLM and LacZ, belonging to different glycoside hydrolase families. The difference in function between them has been unclear so far for practical application. In this study, LacLM and LacZ from L. kefiranofaciens ATCC51647 were cloned into constitutive lactobacillal expression vector pMG36e, respectively. Furtherly, pMG36n-lacs was constructed from pMG36e-lacs by replacing erythromycin with nisin as selective marker for food-grade expressing systems in Lactobacillus plantarum WCFS1, designated recombinant LacLM and LacZ respectively. The results from hydrolysis of o-nitrophenyl-β-galactopyranoside (ONPG) showed that the β-galactosidases activity of the recombinant LacLM and LacZ was 1460% and 670% higher than that of the original L. kefiranofaciens. Moreover, the lactose hydrolytic activity of recombinant LacLM was higher than that of LacZ in milk. Nevertheless, compare to LacZ, in 25% lactose solution the galacto-oligosaccharides (GOS) production of recombinant LacLM was lower. Therefore, two β-galactopyranosides could play different roles in carbohydrate metabolism of L. kefiranofaciens. In addition, the maximal growth rate of two recombinant strains were evaluated with different temperature level and nisin concentration in fermentation assay for practical purpose. The results displayed that 37℃ and 20-40 U/ml nisin were the optimal fermentation conditions for the growth of recombinant β-galactosidase strains. Altogether the food-grade Expression system of recombinant β-galactosidase was feasible for applications in the food and dairy industry.

Isolation and Identification of Korean type Streptococcus mutans (한국형 Streptococcus mutans의 분리 및 동정)

  • 현성희;장성렬;최영길
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.250-258
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    • 1989
  • S. mutans known as a causative causative agent of dental caries was isolated from a carious lesion of Korean in the present study. The physiological, biochemical characteristics and polysaccharide pattern of these isolates were compated with those of four laboratory strains ; AHT(a), FA-1F(b), LM7(e), and OMZ65(g). One hundred strains of oral streptococci were isolated from dental caries sites of Korean (one male and one female). Among these, 3 strains were identified as S. mutans. These strains were able to grow on selective media MS, MST, MSP, MSP1, MSB, MSBT and were stained dark pink when sprayed with solutions of mannitol and TTC. So, these strains were called strain 108, 110, and 120, respectively. Strain 108, 110, and 120 were bacitracin resistnt. As these strains contained particularly hippurate hydrolysis enzyme, they were distinguished from laboratory strains. Apart from laboratory strains, the strain 108 was not capable of fermenting lactose, the strain 110 was not able to ferment sorbitol, inulin, melibiose, raffinose and the strain120 was incapable of fermenting inulin, raffinose. All fractions of extracellular and ecll bound polysaccharide of the strain108, 110, and 120 were consisted of more glucan than fructan. Aside from laboratory strains, the isolated strains were composed of more water-insoluble glucans related adherence on solid surface than water-soluble. According to these results, the strain108, 110, and 120 had native characteristecs of S. mutans, but they were different from laboratory strains in some characteristics. Therefore, each of them was given a name to S. mutans KHC108, KHC110, and KHC120, respcetively.

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Syntheses of Pyrrolo[2,1-b]thiazole Carboxanilides and Their Selective Antifungal Activities against Rice Blast and Wheat Leaf Rust (Pyrrolo[2,1-b]thiazole 카르복스아닐라이드 유도체의 합성 및 그들의 벼 도열병균과 밀 붉은녹병균에 대한 선택적인 항균활성)

  • Hahn, Hoh-Gyu;Nam, Kee-Dal;Yang, Bum-Seung;Choe, Gyeong-Ja;Cho, Kwang-Yun;Lee, Seon-Woo
    • The Korean Journal of Pesticide Science
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    • v.9 no.2
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    • pp.122-131
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    • 2005
  • For development of new agrochemical fungicide, syntheses of pyrrolo[2,1-b]thiazole carboxanilide derivatives 9 and antifungal screening against 6 kinds of plant pathogens were carried out. Functionalization of carboxylic acid on C-2 into carboxanilide in the pyrrolo[2,1-b]thiazole resulted in new 20 candidates, in which are ${\alpha},{\beta}$-unsaturated carboxanilide and methyl groups that are in cis relationship. Treatment of acetoxy-1,4-thiazin with magnesium in refluxing methanol gave pyrrolo[2,1-b]thiazole ethyl ester 4 in higher yield than that of the previous report. Hydrolysis of this compound afforded the corresponding acid 5, which reacted with aniline derivatives in the presence of coupling reagent, DIC to give pyrrolo[2,1-b]thiazole carboxanilides 9. As the result of in vivo antifungal assay of 9 against rice blast, rice sheath blight, cucumber gray mold, tomato late blight, wheat leaf rust, and barley powdery mildew, some compounds showed selectively antifungal activities against the rice blast and wheat leaf rust.

Synthesis of Novel Kojic Acid Derivative and Its Anti-pigmentation Effect (코직산 유도체의 합성과 미백효과)

  • Kim Ki Ho;Kim Ki Soo;Kim Jin Guk;Han Chang Sung;Kim Young Heui;Park Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.3 s.47
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    • pp.409-414
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    • 2004
  • Kojic acid is well known for its anti-pigmentation effect with tyrosinase inhibition activity. However, kojic acid is a unstable compound. In order to improve stability, kojic acid derivative, kojic acid $6-O-2',3',4',6'-tetraacetyl-{\beta}-D-glucopy-ranoside\;(KTGP)$, was synthesized with $O-pentaacetyl-{\beta}-D-glucose$ through the regio- and stereo-selective glycosylation of 6-OH group of kojic acid. High yield $(80\%)$ was obtained by the use of Lewis acid and organic base in nonpolar solvent. Hydrolysis of KTGP with the aid of sodium methoxide in methanol afforded kojic acid $6-O-{\beta}-D-glucopyranoside$ (KGP), which was confirmed by $^1H-NMR\;and\;^{13}C-NMR$ KGP is freely soluble in water and soluble in methanol and ethanol. Inhibition activity of KGP for tyrosinase was investigated by measuring the activity of mushroom tyrosinase compared with those of ascorbic acid, kojic acid, and arbutin. The free radical-scavenger activity was determined by the 1,1-diphenyl- 2-picrylhydrazyl (DPPH) assay. In toxicity assay, KGP was much less toxic than kojic acid and arbutin, Therefore, glycosylation of kojic acid may be useful for the development of stable kojic acid derivatives.

Structure activity relationship on the herbicidal activity by the N-phenyl substituents of 2-(4-(6-chloro-2-benzoxazolyloxy)phenoxy)-N-Phenylpropionamide derivatives in down land (수답에서 2-(4-(6-chloro-2-benz-oxazolyloxy)phenoxy)-N-phenylpropionamide 유도체 중 N-phenyl 치환체들의 제초활성)

  • Sung, Nack-Do;Lee, Sang-Ho;Ko, Young-Kwan;Lee, Kyung-Mo;Kim, Dae-Whang;Kim, Tae-Joon
    • The Korean Journal of Pesticide Science
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    • v.4 no.2
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    • pp.21-28
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    • 2000
  • A new fourty six 2-(4-(6-chloro-2-benzoxazolyloxy)phenoxy)-N-phenylpro- pionamide derivatives were synthesized and the herbicidal activities against rice plant and barnyard grass with pre-emergence in down land were measured. The structure activity relationships (SAR) between the activities and physicochemical parameters of the substituted(X) N-phenyl group in substrates were analyzed and discussed by Free- Wilson and Hansch method from the basis on the former study (Sung. et. al., 1999). The conditions of selective herbicide activity both the barnyard grass and rice plant are shown that the optimal hydrophobicity, $({\pi})_{opt.}=1.34$ and electron donating with field effect (F<0) of meta and ortho, para-substituted mono or disubstituent on the N-phenyl ring were found to contribute significantly. The herbicidal activities against barnyard grass are roughly the same as the results in up land whereas damage to rice plant in down land more increase than that of up land. Degradation products in water are 2-(4-(6-chloro-2-benzoxazolyloxy)phenoxy)propionic acid ((A)) (obs. pKa=4.35 & obs. logP=4.77) and 6-chloro-2-benzoxazolone (B) (obs. pKa=8.40 & obs. logP=2.90). These results were supposing that the hydrolysis product of substrates, (A) is comparatively absorbed in rice plant but not in barnyard grass. And it is assumed from the SAR equations that the 2,6-dimethyl-4-methoxymethyl group substituent ($pI_{50}=5.41$, 3g/ha) is selected as the most highest herbicidal activity against barnyard grass in green house.

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Structural Analysis of the Unusual Sugar-Containing Oligosaccharides Formed by the Selective Cleavage of Weakly Acidic Polysaccharide (약산성 다당의 선택적 분해 과정에서 얻어진 특이당 함유 Oligo당의 구조적 분석)

  • Shin, Kwang-Soon;Lee, Ho
    • Korean Journal of Food Science and Technology
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    • v.29 no.6
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    • pp.1105-1112
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    • 1997
  • By sequential degradation using partial acid hydrolysis of a weakly acidic polysaccharide (GL-4IIb2'), two acidic oligosaccharide fragments, PA-2' and PA-1-III were isolated and their structures were characterized. PA-2' consisted of almost equal proportion of a rhamnose (Rha) and an unusual sugar, 3-deoxy-D-manno-2-octurosonic acid (Kdo). When permethylated oligosaccharide-alditol derived from PA-2' was analyzed by GC-MS, the peak gave the fragment ions at m/z 189 $(bA_1,\;6-deoxyhexose)$ and at m/z 308 $(aJ_2,\;alditol\;from\;Kdo)$. The peak also gave the characteristic ion at m/z 162 but it did not give the fragment ion at m/z 177, suggesting that Kdo is substituted at C5 but not at C4. Methylation analysis also indicated that PA-2' was composed mainly of terminal Rhap and 5-substituted Kdo. When the reduced product from PA-2' was analyzed by $^1H-NMR$, it gave a signal at 5.09 ppm due to an anomeric proton of ${\alpha}-L-Rha$. These results indicated that PA-2' mainly contained ${\alpha}-L-Rhap-(1{\rightarrow}5)-Kdo$. On the other hand, PA-1-III mainly comprised Rha and Kdo in addition to small proportions of arabinose (Ara) and 3-deoxy-D-lyxo-2-heptulosaric acid (Dha). MS analysis of permethylated oligosaccharide-alditols from PA-1-III suggested that the major peak 1P was $Rhap-(1{\rightarrow}5)-Kdo$ whereas the minor peaks 2P and 3P possessed $Araf-(1{\rightarrow}5)-Dha$ unit and these peaks were produced as epimers during reduction of carbonyl groups in Dha.

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