• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1580-1586
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• 2007

The objective of this study was to compare the bioefficacy of L-lysine sulfate relative to L-lysine${\cdot}$HCl for 10 to 20 kg pigs. Two experiments were conducted to determine the bioefficacy of the two sources of lysine using daily gain, feed conversion, plasma urea nitrogen and nitrogen retention as the response criteria. In experiment 1, 168 crossbred barrows ($Landrace{\times}Large$ White), weaned at $28{\pm}3$ d ($9.07{\pm}0.78$kg body weight), were allotted to one of seven dietary treatments in a $2{\times}3$ (two lysine $sources{\times}three $ lysine levels) factorial arrangement of treatments with an added negative control treatment group. The basal diet was based on corn, peanut meal and soybean meal and provided 0.67% lysine. The basal diet was supplemented with 0.1, 0.2 or 0.3% lysine equivalents supplied from either L-lysine sulfate or L-lysine${\cdot}$HCl. Each treatment was fed to six pens of pigs with four pigs per pen. The trial lasted 21 days. The relative bioefficacy value of lysine in L-lysine sulfate using daily gain, feed conversion and plasma urea nitrogen as response criteria was 1.01, 1.05 and 1.04 of the lysine in L-lysine${\cdot}$HCl, respectively. In experiment 2, 42 crossbred ($Landrace{\times}Large$ White) pigs ($16.03{\pm}1.58$ kg body weight) were housed in stainless steel metabolism cages for 10 d and fed the seven diets used in the nitrogen-balance trial. The relative bioefficacy value of L-lysine sulfate was estimated to be 0.95 as effective as L-lysine${\cdot}$HCl for nitrogen retention on an equimolar basis. The t-test analysis revealed that bioefficacy of lysine in L-lysine sulfate was not significantly different from lysine in L-lysine${\cdot}$HCl, which was set at 1.00. In conclusion, L-lysine sulfate can be used instead of L-lysine${\cdot}$HCl to fortify lysine-deficient diets fed to 10 to 20 kg pigs.

• KSBB Journal
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• v.16 no.5
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• pp.474-479
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• 2001

Fed-batch culture, single stage and two stage continuous cultures of Corynebacterium glutamicum SH 35 for the production of $_{L}$-Iysine were performed and compared. In the case of fed batch culture, $_{L}$-Iysine concentration, $_{L}$-Iysine yield and $_{L}$-Iysine productivity was 129.2 g/L, 47.0% and 3.08 g/L/h, respectively. In a single-stage continuous culture, optimum dilution rate and pH was 0.1 h$^{-1}$ and 6.9, respectively, and optimum concentration of sugar and ammonium sulfate in a medium reservoir was 108 g/L and 25 g/L, respectively. Under the optimized conditions, 67 of cell concentration($OD_{610}$), 44.2 g/L of lysine concentration, 41% of $_{L}$-Iysine yield and 4.39 g$L^{-1}$ of $_{L}$-Iysine productivity were obtained. In a two-stage continuous culture, optimum dilution rate was 0.075 $h^{-1}$. Under the conditions, 103 of cell concentration($OD_{610}$) 84.0g/L of $_{L}$-Iysine concentration and 46% of $_{L}$-Iysine yield were obtained.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.375-380
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• 1987

D-$\alpha$-Amino-$\varepsilon$-carpolactam racemase (EC 5.1.1) and L-$\alpha$-amino-$\varepsilon$-caprolactam hydrolase (EC 3.5.2) were fractionated from cell-free extracts of Alcaligenes eutrophus A52 using ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. It was made sure that D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-$\alpha$-amino-$\varepsilon$-caprolactam by racemase, and then hydrolyzed into L-lysine by hydrolase in Alcaligenes eutrophus A52. For the conversion of D-$\alpha$-amino-$\varepsilon$-caprolactam into L-lysine by cell-free extracts of Alcaligenes eutrophus A52, the optimum temperature and pH were 6$0^{\circ}C$ and 8.5 respectively. The results showed that 0.5% D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-lysine at 55$^{\circ}C$ for 10 hr with a conversion rate of 98% by cell-free extracts containing 3.1mg of protein.

• Journal of the Korean Society of Food Science and Nutrition
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• v.16 no.4
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• pp.251-261
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• 1987

This study was attempted to increase the production of L-Threonine by Brevibacterium Flavum ATCC 14067, To select the strain which produce the highest threonine, mutants ere induced by N-methyl-N'-nitro-N-nitrosoguanidine treatment. The composition of media and cultural condition for its overproduction of threonine were also studied. In a threonine producer, strain B-13(Met-) was the strain producing the highest amount of threonige among methionine, lysine and isoleucine auxotrophs. The following results were obtained. 1. The wild strain and B-13(Met-) produced threonine 1.4mg/ml and 4.86mg/ml , respectively. 2. The optimum composition of medium for producing threonine by Brevibacterium Flavum B-13 was glucose 10%, ammonium sulfate 4%, potassium phosphate monobasic 0.2%, magnesium sulfate 0.05%, biotin $200{\mu}l$, thiamine $300{\mu}l$. Addition of nicotinic acid also led to increase L-threonine production. 3. In addition of organic nutrients to the fermentation medium, peptone n'ere effective and addition of methionine $100{\mu}g/ml$ produced the highest amount of L-Threonine. Aspartic acid and homoserine were also effective when these amino acid were added to the fermentstion medium. 4. Cultural conditon on threonine production by B-16 were investigated. The optimum pH was 7.0-8.0. The highest amount of threnine was produced after 4 days of cultural period.

• 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 Korean Journal of Mycology
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• v.14 no.2
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• pp.101-107
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• 1986

The properties of carboxyl proteinase which was contained in Poria cocos (Schw.) Wolf were investigated by means of the purification with 0.65 ammonium sulfate saturation, DEAE cellulose and Sephadex G-75 gel filtration. This enzyme was found to hydrolyze only peptide bond between glutamyl-L-tyrosine of carbobenzoxy-L-glutamyl-L-tyrosine among the synthetic substrates of carbobenzoxy-L-glutamyl-L-tyrosine, hippuryl- L-phenylalanine and hippuryl-L-arginine. This enzyme was inhibited by $Zn^{+2},\;Fe^{+2},\;Ca^{+2},\;CN^{-1},\;P_2O_7^{-4}$ ions, but stimulated by $Hg^{+2}$ ion. Also, this enzyme was inhibited by organic compounds such as L-lysine, L-phenylalanine, hippuryl-L-phenylalanine, diazoacetyl-DL-norleucine methyl ester (DAN) and 1,2-epoxy-3-(P-nitrophenoxy)propane(EPNP). In particular, the activity was inhibited by L-lysine till 20 minutes of preincubation time rapidly, and by DAN in the presence of $Cu^{+2}$ ion more rapidly after 30 minutes than DAN in the absence of $Cu^{+2}$ ion. L-Lysine was found to be a competitive inhibitor and its $K_i$ value was determined to be 0.12 mmole by Dixon plot.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.226-232
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• 2016

Dihydrodipicolinate reductase is an enzyme that converts dihydrodipicolinate to tetrahydrodipicolinate using an NAD(P)H cofactor in L-lysine biosynthesis. To increase the understanding of the molecular mechanisms of lysine biosynthesis, we determined the crystal structure of dihydrodipicolinate reductase from Corynebacterium glutamicum (CgDapB). CgDapB functions as a tetramer, and each protomer is composed of two domains, an Nterminal domain and a C-terminal domain. The N-terminal domain mainly contributes to nucleotide binding, whereas the C-terminal domain is involved in substrate binding. We elucidated the mode of cofactor binding to CgDapB by determining the crystal structure of the enzyme in complex with NADP⁺ and found that CgDapB utilizes both NADH and NADPH as cofactors. Moreover, we determined the substrate binding mode of the enzyme based on the coordination mode of two sulfate ions in our structure. Compared with Mycobacterium tuberculosis DapB in complex with its cofactor and inhibitor, we propose that the domain movement for active site constitution occurs when both cofactor and substrate bind to the enzyme.

• Journal of Plant Biotechnology
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• v.3 no.3
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• pp.151-157
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• 2001

In order to breed barley lines with reduced viscosity of wort, a fractured starch mutant of naked barley cultivar, Nubet, was obtained from the M2 seeds mutated by the diethyl sulfate treatment. Seeds of this fractured starch mutant were opaque and the endosperm consists of angular, irregular and fractured starch. The mutant was caused by single recessive mutation and assigned by the symbol fra. The gene was located on chromosome 4, distal in long arm by linkage recombinations using translocation homozygote lethal test set. The linkage value between the fractured starch mutant and 72-4a, 72-4d were 26.0$\pm$4.9, 34.2$\pm$3.1 percent respectively. In addition to the reduced seed size, fewer kernels per spike and higher tillering ability, lower $\beta$-glucan viscosity and higher lysine content of the grain were associated with this mutant. $\beta$-glucan viscosity of the Nubet grains increased from 3 weeks after anthesis to matury and most of the viscose substances appeared to be stored in the middle of the endosperm tissue. Since the mutant grains showed better milling property as compared to Nubet, it can be used as breeding resources to develope new barley cultivars for maltins and milling purpose.

• Fisheries and Aquatic Sciences
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• v.7 no.3
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• pp.99-108
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• 2004

Vitellogenin (VTG) was purified from serum of $estradiol-l7{\beta}-treated rockfish$(Sebastes schlegeli) by precipitation with $EDTA-Mg^{2+}$ and ammonium sulfate and two step chromatography (anion exchange chromatography and gel permeation chromatography) was performed on FPLC system. Rockfish VTG (rfVTG) was characterized and its properties were determined. The monomers have apparent, molecular mass of about 188 kDa as indicated by SDS-PAGE. Amino acid composition analysis of rfVTG was similar to VTG from other oviparous teleosts. Cysteine and lysine were present at relatively high level. Leucine was present at relatively lower level than in other species. The N-terminal amino acid sequence was evaluated to identify rfVTG. Western blot analysis using an antibody against the purified VTG showed that the antibody reacted with both plasma of $estradiol-l7{\beta}-treated rockfish$ treated male and purified VTG, whereas there was no reaction with male serum of the control. An ELISA was developed using monoclonal and polyclonal antibodies against rfVTG. The assay range was 3.2 ng/mL and 1,000 ng/mL and the value of the intra and inter assay variations were within $9.7{\%}$ and $11.2{\%}$, respectively. Recovery rate was $96.8{\%}$. The sandwich ELISA could be useful for the detection of VTG and could be good for screening of estrogenic compounds.

• Fisheries and Aquatic Sciences
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• v.2 no.2
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• pp.218-225
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• 1999

A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.