• Title/Summary/Keyword: L-Lysine Production

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Improved Poly-${\varepsilon}$-Lysine Biosynthesis Using Streptomyces noursei NRRL 5126 by Controlling Dissolved Oxygen During Fermentation

  • Bankar, Sandip B.;Singhal, Rekha S.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.652-658
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    • 2011
  • The growth kinetics of Streptomyces noursei NRRL 5126 was investigated under different aeration and agitation combinations in a 5.0 l stirred tank fermenter. Poly-${\varepsilon}$-lysine biosynthesis, cell mass formation, and glycerol utilization rates were affected markedly by both aeration and agitation. An agitation speed of 300 rpm and aeration rate at 2.0 vvm supported better yields of 1,622.81 mg/l with highest specific productivity of 15 mg/l.h. Fermentation kinetics performed under different aeration and agitation conditions showed poly- ${\varepsilon}$-lysine fermentation to be a growth-associated production. A constant DO at 40% in the growth phase and 20% in the production phase increased the poly-${\varepsilon}$-lysine yield as well as cell mass to their maximum values of 1,992.35 mg/l and 20.73 g/l, respectively. The oxygen transfer rate (OTR), oxygen utilization rate (OUR), and specific oxygen uptake rates ($qO_2$) in the fermentation broth increased in the growth phase and remained unchanged in the stationary phase.

Continuous Fermentationof L-Lysine by Immobilized Corynebacterium glutamicum (Corynebacterium glutamicum 고정화균체에 의한 L-라이신 연속발효)

  • 이인선;조정일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.2
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    • pp.322-327
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    • 1994
  • For the improvement of L-lysine productivity, development of the continuous fermentation system by a bioreactor assembly was attempted. Primarily, optimal conditions on the whole cell immobilization of Corynebacterium glutamicum ATCC21514 were studied and 76.2% of immobilization ratio was obtained when the cells were entrapped with 4% k-carrageenan showing 4.0kg gel strength. A bioreactor system was set up using the immobilized cells was applied for the continuous production of L-lysine. The results obtained under the optimum conditions were compared with those of the batchwise fermentation. Experimental results obtained from 14 day continuous fermentation showed 36.7% of sugar conversion to L-lysine while the productivity of L-lysine was disclosed as 4.96mg/ml mg-dry cell weight /hr which is 2.5times and 4.1 times higher than those of the batchwise fermentation by the intact cells and by the immobilized cells, respectively.

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Enhancement of Lysine Production in Recombinant Corynebacterium glutamicum through Expression of Deinococcus radiodurans pprM and dr1558 Genes (Deinococcus radiodurans 유래 DR1558과 PprM에 의한 Corynebacterium glutamicum의 라이신 생산 향상 연구)

  • Kim, Su-mi;Lim, Sangyong;Park, Si Jae;Joo, Jeong Chan;Choi, Jong-il
    • Microbiology and Biotechnology Letters
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    • v.45 no.3
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    • pp.271-275
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    • 2017
  • The expression of Deinococcus radiodurans dr1558 and pprM genes was examined for enhanced lysine production in recombinant Corynebacterium glutamicum. These genes are known to confer high tolerance to pH and osmotic shock in Escherichia coli. D. radiodurans dr1558 and pprM genes were expressed in C. glutamicum by using 6 synthetic promoters of different strengths, to evaluate the effect of expression efficiency on lysine production. Recombinant C. glutamicum expressing DR1558 under the L26 and I64 promoters showed higher lysine production than that expressing DR1558 under other promoters. Similarly, recombinant C. glutamicum expressing PprM under same promoters (L26 and I64) showed a higher increase in lysine production compared to that expressing PprM under other promoters. In the absence of $CaCO_3$ in the medium, the expression of DR1558 or PprM also increased lysine concentration in C. glutamicum depending on the promoter used. Together, these results suggest that genes involved in radiation tolerance in D. radiodurans can be used to enhance production of amino acids and their derivatives.

Site-speci fic Inactivation o meso-Diaminopimelate-dehydrogenase Gene (ddh) in a Lysine-producing Brevibacterium lactofementum. (Brevibacterium lactofermentum 에서 meso-Diaminopimelate-dehydrogenase Gene (ddh)의 Site-specific Inactivation)

  • 김옥미;박선희;이갑랑
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.387-392
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    • 1998
  • Brevibacterium lactofermentum, a gram-positive bacteria, has both the diaminopimelate (DAP) pathway and meso-DAP-dehydrogenase (DDH) pathway for L-lysine biosynthesis. To investigate importance of DDH pathway and the related ddh gene in lysine production, we introduced site-specific mutagenesis technique. A 300 bp DNA fragment central to the meso-DAP-dehydrogenase gene (ddh) of B. lactofermentum was used to inactive chromosomal ddh gene via homologous recombination. Southern hybridization analysis confirmed that the chromosomal ddh gene was disrupted by the vector sequence. The B. lactofementum ddh mutant obtained have an inactive DDH pathway. The results reveal that inactivation of the ddh gene in B. lactofermentum leads to dramatic reduction of lysine production as well as decrease of the growth rate, indicating that the DDH pathway is essential for high-level lysine production as well as biosynthesis of meso-DAP.

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Development of L-Lysine Producing Strains from Cellulosic Substrate by the Intergeneric Protoplast Fusion - Conditions for Fusion and Properties of Fusants- (속간 원형질체 융합에 의한 섬유질 기질로부터 L-Lysine 생산균주 개발 -융합조건 및 융합체의 성질 -)

  • 성낙계;정덕화;박법규;정영철;전효곤
    • Microbiology and Biotechnology Letters
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    • v.16 no.3
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    • pp.175-181
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    • 1988
  • To produce L-lysine from cellulosic substrate, the intergeneric protoplast fusion between Cellulomonas flavigena and Corynebacterium glutamicum, Cellulomonas flavigena and Brevibacterium flavum was performed. The fusion frequencies were 1.9$\times$10$^{-6}$ to 2.1$\times$10$^{-6}$ for the regenerated protoplasts when two parental strains were treated with 30% of polyethyleneglycol (M.W.6000) containing 5 mM EDTA at 3$0^{\circ}C$ for 30 min. Two fusants, FCB3 and FCC 19 were finally selected by comparision of their genetic stability and L-lysine productivity. The properties of fusants-DNA con-tent, G+C content and L-lysine productivity-were investigated. The DNA content of fusants was greater than those of the parental strain and their G+C contents are equal to half of total G+C con-tent of two parental strains. The fusants showed high productivity of L-lysine from carboxy methyl cellulose as substrate.

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Effects of L-Carnitine with Different Lysine Levels on Growth and Nutrient Digestibility in Pigs Weaned at 21 Days of Age

  • Cho, W.T.;Kim, J.H.;Han, In K.;Han, Y.K.;Heo, K.N.;Odle, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.5
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    • pp.799-805
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    • 1999
  • This study was conducted to investigate the effects of L-carnitine with different levels of lysine on performance of pigs weaned at 21 days of age. A total of 120 pigs were allotted into a $3{\times}2$ factorial design with three different levels of lysine (1.40%, 1,60% and 1.80%) and two levels of L-carnitine (0 and 1,000 ppm). Each treatment had 4 replications with 5 pigs per replicate. Pigs of $22{\pm}1$ days (5.9 kg of body weight) were grouped into a completely randomized block design. Treatments were 1) 1.4-Crt; 1.40% of lysine with 1,000 ppm of L-carnitine, 2) 1.4-N; 1.40% of lysine without L-carnitine, 3) 1.6-Crt; 1.60% of lysine with 1,000 ppm of L-carnitine, 4) 1.6-N; 1.60% of lysine without L-carnitine, 5) 1.8-Crt; 1.80% of lysine with 1,000 ppm of L-carnitine and 6) 1.8-N; 1.80% of lysine without L-carnitine. Growth performance was optimized in pigs fed 1.6% lysine regardless of carnitine addition. For the first 7 days of the experimental period, the best ADG and F/G were found in pigs within the 1.6-Crt group. Carnitine significantly improved (p<0.05) ADG of pigs when the lysine level in the diet was 1.6%. Only in the third week carnitine had a significant influence on growth performance of pigs. A lysine-sparing effect of L-carnitine was not detected in this study. The 1.6-Crt group showed the best proximate nutrient digestibility, and the crude fat and gross energy digestibility were higher when the L-carnitine was added in the diet. Lysine level significantly affected the digestibilities of DM (p<0.001), GE (p<0.001), CP (p<0.01) and C.fat (p<0.05). Carnitine also significantly improved digestibility of nutrients. Lysine level as well as carnitine level affected the amino acids digestibility, however, in 1.8% lysine diet carnitine did not influence on amino acids digestibility. Plasma carnitine content was significant higher (p<0.05) in pigs fed L-carnitine. This indicates the increased biological availability of carnitine within the body. L-carnitine supplementation tended to improve feed utilization during the third week (p<0.10) and during the entire period (p=0.10). Lysine level significantly affected feed utilization of pigs during the third week and entire period (p<0.05). As pigs grew, the lysine requirement was reduced.

L-Lysine Production by 6-Azauracil Resistant Mutant of Corynebacterium glutamicum (6-Azaumcil 내성을 지닌 Corynebacterium glutamicum 변이주에 의한 L-Lysine의 생산)

  • 신현철;김성준전영중이재흥
    • KSBB Journal
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    • v.9 no.4
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    • pp.372-377
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    • 1994
  • To improve L-lysine yield, pyrimidine base analogue(6-azauracil)-resistant mutants were isolated from Corynebacterium glutamicum KFCC10672 Among them the best producer, C. glutamicum CH0516, was selected and tested for L-lysine production in a $7\ell$ fermentor. It was found that the product yield obtained with C. glutamicum CH0516 was higher than that of the parent strain by 3%. In order to elucidate the gain in productivity with the 6-azauracil-resistant mutant enzymatic kinetic parameters such as aspartokinase(AKase) and aspartate carbamoyltransferase (ATCase) were measured. The Km values of AKase with C. glutamicum KFCC10672 and CH0516 were 200.0 mM and 166.7 mM and those of ATCase were 0.13 mM and 0.27 mM, respectively. However, the specific enzyme activities of AKase of C. glutamlcum KFCC10672 and CH0516 were $3.89{\times}10^{-1}$ units/mg and $4.78{\times}10^{-1}$ units/mg, and those of ATCarse were 2.20 units/mg and 1.84 units/mg, respectively. It appears that some increase in product yield with C. gluramicum CH0516 is likely due to the increased Akase activity and decreased ATCase activity.

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Effects of Dietary Metabolizable Energy and Lysine on CarcassCharacteristics and Meat Quality in Arbor Acres Broilers

  • Tang, M.Y.;Ma, Q.G.;Chen, X.D.;Ji, C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.12
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    • pp.1865-1873
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    • 2007
  • An experiment was performed to evaluate the effects of dietary metabolizable energy (ME) and lysine on carcass characteristics and meat quality in Arbor Acres (AA) broilers from 1 to 56 days of age. A total of 2,970 1-d-old male broiler chicks were randomly allocated to nine dietary treatments (three ME levels in combination with three lysine levels), and dietary ME and lysine concentrations were formulated by varying corn, soybean meal, tallow, and L-lysine sulfate concentrations. Live body weight (BW), carcass weight (CW), dressing percent, breast muscle weight (BMW), yield of breast muscle, muscle color (CIE L*, a*, and b*), pH values 45 min and 24 h postmortem ($pH_{45}$, and $pH_{24}$), meat shear force value (SFV), and water loss rate (WLR) were evaluated. Results showed that live body weight and dressing percent increased (p<0.05) as dietary energy increased. Higher dietary lysine content improved breast muscle weight. Neither carcass weight nor yield of breast muscle was affected by dietary energy or lysine content. Higher ME increased the b* value (p = 0.067) and $pH_{24}$ value (p<0.05), whereas it decreased SFV (p<0.05) and WLR (p = 0.06). Only water loss rate was influenced (p<0.01) by dietary lysine, which was higher in broilers from the high lysine diet as compared to those from medium or low lysine diets. The $pH_{45}$ value and L* value of breast muscle were not affected by ME or lysine. Significant interaction of dietary ME and lysine was found on a* value of breast muscle. These results indicated that dietary ME and lysine had important effects on breast muscle growth and meat quality, however their effects were different. Different concentrations of dietary ME and lysine might be considered to improve meat quality.

The Effect of Redox Potential on the Kinetics of Lysine Production by Corynebacterium glutamicum (Corynebacterium glutamicum에 의한 Lysine 생산에 있어서 산화환원 전위가 발효속도론적 특성에 미치는 영향)

  • 이진희;김성준;이재흥
    • Microbiology and Biotechnology Letters
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    • v.19 no.1
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    • pp.76-81
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    • 1991
  • - The effect of redox potential (ORP) on lysine production by a leucine auxotrophic regulatory mutant of Corynebacterium glutclmicum on molasses medium was investigated in a 2-1 jar fermentor at pH 6.9 and $32^{\circ}C$. At a dilution rate of D=O.l $h ^1$, a maximum yield of Yr,,s=0.24 was obtained in either carbon- or leucine-limited chemostat where the redox potential was between -60 mV and - 100 mV. This level of redox potential corresponded to moderate oxygen deficiency. Under a high oxygen deficient condition of the redox potential of - 130 rnV (oxygen-limited chemostat), all the kinetic parameters such as $Y_[p/s}, q_s\; and \; q_p$ were decreased significantly and significant amounts of byproducts including glycine, alanine and valine were accumulated in the culture, indicating that the control of redox potential is important in lysine fermentation. At the redox potential of - 40 mV, on the other hand, large quantities of arginine (up to 0.38g/l) and glutamic acid (up to 0.12 g/l) were produced. A maximum lysine productivity of 2.41 g/l/h was achieved at - 66 mV under a carbon-limited condition.

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Nutritional Regulation of GLUT Expression, Glucose Metabolism, and Intramuscular Fat Content in Porcine Muscle

  • Katsumata, M.;Kaji, Y.;Takada, R.;Dauncey, M.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.8
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    • pp.1297-1304
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    • 2007
  • We conducted a series of investigations in order to elucidate role of nutritional status in regulating GLUT expression and energy metabolism in porcine muscle. Firstly, the role of mild undernutrition in regulating muscle GLUT gene expression and function was studied in growing pigs (3 wk of age) on a high (H) or low (L) food intake (H = 2L) at $35^{\circ}C$ or $26^{\circ}C$. Low food intake selectively upregulates GLUT1 and GLUT4 gene expression; mRNA levels were elevated in longissimus dorsi (L. dorsi) and rhomboideus muscles but not in diaphragm or cardiac muscles. Our next step was to determine whether dietary lysine, a major primary limiting amino acid in diets for pigs, affects muscle GLUT4 expression. Pigs of 6 wk of age were pair-fed a control or low lysine (LL) diet. The control diet contained optimal amounts of all essential amino acids, including 1.15% lysine. The LL diet was similar but contained only 0.70% lysine. GLUT4 mRNA expression was upregulated by the LL diet in L. dorsi and rhomboideus muscles, whereas that in cardiac muscle was unaffected. GLUT4 protein abundance was also higher in rhomboideus muscle of animals on the LL diet. We conducted another investigation in order to elucidate effects of the LL diet on post-GLUT4 glucose metabolism. Activity of hexokinase was unaffected by dietary lysine levels while that of citrate synthase was higher both in L. dorsi and rhomboideus muscles of pigs fed on the LL diet. Glucose 6-phosphate content was higher in L. dorsi msucle in the LL group. Glycogen content was higher both in L. dorsi and rhomboideus muscles in the LL group. Further, we determined the effects of dietary lysine levels on accumulation of intramuscular fat (IMF) in L. dorsi muscle of finishing pigs. A low lysine diet (lysine content was 0.40%) meeting approximately 70% of the requirement of lysine was given to finishing pigs for two months. IMF contents in L. dorsi of the pigs given the low lysine diet were twice higher than those of the pigs fed on a control diet (lysine content was 0.65%). Finally, we proved that a well known effect of breadcrumbs feeding to enhance IMF of finishing pigs could be attributed to shortage of amino acids in diets including breadcrumbs.