• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.4
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• pp.336-341
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• 2002

This experiment was conducted to evaluate the lysine cell mass (LCM) as a dietary fish meal (EM) protein replacer in juvenile Israeli carp, Cyprinus carpio. Fishmeal, a major animal protein source in the control diet, was replaced by tCM on the protein equivalent base, Fish averaging 1,7 $\pm$ 0.1 g (Mean $\pm$ SD) fed one of nine diets containing isonitrogenous and isocaloric basis of $38\%$ crude protein and 15.2 kJ available energy/g diet: control, $100\%$ $FM; LCM_20$, $80\%$ $FM+20\%$ $LCM; LCM_40$, $60\%$ $FM+40\%$ $LCM; LCM_60$, $40\%$ $FM+60\%$ $LCM; LCM_100$, $100\%$ $LCM; LCM_20$l, $80\%$ $FM+20\%$ $LCM+0.07\%$ $Lysine; LCM_40$l, $60\%$ $FM+40\%$ $LCM+0.14\%$ $Lysine; LCM_60$l $40\%$ $FM+60\%$ $LCM+0.22\%$ Lysine; LCM_100l, $100\%$ LCM+$0.35\% Lysine. After 6 weeks of feeding trial there was no significant difference in weight gain (WG), feed efficiency (FE), protein efficiency ratio (PER) and specific growth rate (SGR) among fish fed control and $LCM_20$ (P>0.05), while fish fed $LCM_40,\;LCM_60,\;LCM_100,\;LCM_40l,\;LCM_60l\;and\;LCM_100l$ diets had a significantly lower WG, FE, PER and SGR than did fish fed control diet (P<0.05). There was no significant difference in WG, PER and SGR among fish fed control and $LCM_20$l diets (P>0.05), while fish fed $LCM_20$l S had a significantly lower FE than did fish fed control diet (P<0.05). No significant difference was observed in hematocrit and condition facto, among fish fed nine diets (P>0.05). Therefore, these results indicated that LCM could replace FM up to $20\%$ and dietary synthetic lysine supplementation did not show any positive growth effects in juvenile Israeli carp.

• Journal of Aquaculture
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• v.17 no.2
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• pp.122-127
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• 2004

On protein equivalence base, fishmeal (FM) was replaced by lysine cell mass (LCM) in selected different diets in Korean rockfish, Sebastes schlegeli (Hilgendorf) Eight experimental diets were formulated to contain 100% FM (LC $M_{0}$), 90% FM+10% LCM (LC $M_{10}$),80% FM+20% LCM (LC $M_{20}$), 70% FM+30% LCM (LC $M_{30}$), 60% FM+40% LCM (LC $M_{40}$ ), 70% FH+30% LCM+lysine (LC $M_{+Lys}$), 60% FM+40% LCM+lysine (LC $M_{40+Lys}$), and 50% FM+50% LCM+lysine (LC $M_{50+Lys}$). Experimental individuals of the fish (12.6 g) were randomly fed on one of the experimental diets. After 6 weeks of feeding trial, weight gain (WG) and feed efficiency (FE) of fish fed LC $M_{0}$ diet was significantly (P〈0.05) higher than those of fish fed LC $M_{20}$, LC $M_{30}$, LC $M_{40}$ , LC $M_{30+Lys}$, LC $M_{40+Lys}$, and LC $M_{50+Lys}$ diets, however, there was no significant difference in WG of fish fed LC $M_{0}$ and LC $M_{10}$ diets. Supplementation of lysine has no effect on WG. There was no significant difference in condition factor (CF) of fish fed LC $M_{0}$, LC $M_{10}$ and LC $M_{20}$ diets. Hemoglobin (Hb) of fish fed LC $M_{0}$, LC $M_{10}$, LC $M_{20}$, LC $M_{30}$, LC $M_{40}$ , LC $M_{30+Lys}$, and LC $M_{40+Lys}$, diets were not significantly different from each other. No significant differences were observed in hematocrit (PCV) and hepatosomatic index (HSI) among all dietary treatments. Apparent digestibility of dry matter (ADM) and protein (ADP) of diets significantly decreased with increase in dietary LCM level, though there was no difference in ADM and ADP between LC $M_{0}$ and LC $M_{10}$. These results indicate that LCM could replace up to 10% of fishmeal in Korean rockfish diets.ish diets.iets.ish diets.s.ish diets.

• Journal of Aquaculture
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• v.14 no.3
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• pp.197-203
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• 2001

To replace fish meal (FM) in the diet of Nile tilapia, Oreochromis nloticus, different levels of Iysine cell mass (LCM) were added to diet on protein equivalent base. fish averaging 3.0 g fed one of nine diets containing isonitrogenous and isocaloric basis of 35% crude protein and 15.3kJ available energy/g diet: $LCM_0, 100% FM: LCM_{10}, 90% FM+10% LCM, LCM_{20}, 80% FM+20% LCM; LCM_{30}, 70% FM+30% LCM; LCM_{40}, 60% FM+40% LCM; LCM_{30}l, 70% FM+30% LCM+lysine; LCM_{30}ln, 70% FM +30% LCM + lysine +NaOH; LCM_{40}lan, 60% FM+40% LCM+lysine+arginine; LCM_{40}/lan, 60% FM+40% LCM+lysine+arginine+NaOH.$. After 8 weeks of feeding trial, there were no significant differences in weight gain (WG) and specific growth rate (SGR) among fish fed LCM$_{0}$ (control diet), LCM_{10}, LCM_{30}, LCM_{40}, LCM_{30}l, LCM_{30}ln, LCA_{40}la and LCM_{40}lan diets (P>0.05)$, while fishes fed $LCM_{20} diet were significantly higher than those fed LCM_{0}, LCM_{30}, LCM_{40}, LCM_{40}la and LCM_{40$}lan diets (P<0.05). There were no significant differences in feed efficiency (FE) and protein efficiency ratio (PER) among fish fed control diet, $LCM_{10}, LCM_{20}, LCM_{30}ln, LCM_{ 40}la and LCM_{40}$lan diets (P>0.05), while fishes fed control diet were significantly higher than those fed $LCM_{30}, LCM_{40} and LCM_{40}l diets (P<0.05)$. Positive effects were not shown in WG and SGR with supplementation of amino acids (lysine & arginine) and neutralizatio, while FE and PER from fish fed $LCM_{40}la and LCM_{40}lan diets were significantly higher than those fed LCM_{30}, LCM_{40} and LCM_{30}$l diets (P<0.05). Hence, LCM can replace FM up to 40%, and dietary suppl-ementacon and neutralization of amino acids showed positive effects, when FE and PER were considered in juvenile tilapia diet.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.5
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• pp.550-558
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• 1998

The experiment was conducted to evaluate CM (Cell Mass from Lysine Fermentation), which is used to produce synthetic lysine in industry, as an alternative protein source in broiler diets. Three different production conditions were employed to produce CMs (CM I, II, III). Treatments were control, CM I -1 (1 % of CM in the diet), CM I -3 (3% of CM in the diet), CM I -5 (5% of CM in the diet), CM II (3% of CM in the diet), and CM III (3% of CM in the diet). It was found that CM products were all high in crude protein content and especially high in lysine and methionine contents, while very low in minerals. For the starter period, all CM groups showed better weight gain, chicks fed CM I -1 diets were especially high in weight gain (p < 0.05). CM groups consumed 14.4 to 18.0% more feed than chicks fed control diets (p < 0.05). The best FCR was found in CM I -1 groups (p < 0.05), but as CM level was increased, FCR was also increased. For the finisher period, weight gain was similar through all treatments. Through whole experimental period, weight gain and feed intake were higher in all CM groups than control group (p < 0.05), however, as CM level was increased, FCR was also increased. Generally chicks fed CM diets showed higher utilizabilities of gross energy, dry matter, crude protein and crude fat. The best nutrients utilizability was obtained in CM I -1 group, and the worst was found in the control group. During the finisher period, the utilizabilities of crude protein, crude ash and phosphorus were not affected by the dietary treatments. Amino acids utilizability was not significantly affected by the treatments except CM I -5 group. In all amino acids tested, chicks did not show the big difference in utilizabilities. Only in the CM I -5 group, amino acids utilizability was significantly lower than control group. However, among CM I groups, the mean value of the amino acids utilizability was decreased as the level of CM inclusion in the diet was increased. During the finisher period, similar trend was found in amino acids utilizability.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.556-562
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• 1989

The bioconversion of mandarin orange peel press liquor to single cell protein (SCP) by two yeast strains, F-60, and C-7, which were isolated from mandarin orange peel was carried out and compared with that of using Candida utilis IFO 0598. Experiments were directed toward the high yield of biomass and high protein in cultures of the strains mentioned above. Candida utilis IFO 0598, F-60 and C-7 strains were cultivated at 3$0^{\circ}C$, pH 5.2 for 3 days in shaking flasks. The effects of some nutrients on cell growth were studied. Cell mass and protein content per cell mass were increased by addition of urea 1%, KH$_2$PO$_4$ 0.1% and MgSO$_4$ㆍ7$H_2O$ 0.05%, When the F-60 strain cultured under the optimal conditions, cell mass, growth yield and protein content were 41.2g/l, 53.9%, 59.7%, respectively. Cell mass was also increased up to 15% by modifying the fermentation condition on the bench type 20l jar fermentor. Crude fat content (10.3%) of dried C-7 cell was higher than those of C. utilis and F-60, 4.9% and 5.6% respectively. Total protein content of the F-60 strain was 59.7% per dry weight. And we compared their amino acid compositions with that of FAO provisional pattern. In the case of the F-60 strains, amino acid contents such as lysine, leucine and isoleucine were much higher than those of methionine, cystine and tryptophan.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.364-370
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• 2008

Chemical analysis and in vitro studies were conducted to investigate the nutritive value for ruminants of cell mass from lysine production (CMLP) which is a by-product of the lysine manufacturing process. Proximate analysis, protein fractionation, and in vitro protein degradation using protease from Streptomyces griseus and strained ruminal fluid were carried out to estimate ruminal protein degradability of CMLP with two reference feedstuffs-soybean meal (SBM) and fish meal (FM). Amino acid composition and pepsin-HCl degradability were also determined to evaluate postruminal availability. CMLP contained 67.8% crude protein with a major portion being soluble form (45.4% CP) which was composed of mainly ammonium nitrogen (81.8% soluble CP). The amount of nucleic acids was low (1.15% DM). The total amount of amino acids contained in CMLP was 40.60% DM, which was lower than SBM (47.69% DM) or FM (54.08% DM). CMLP was composed of mainly fraction A and fraction B2, while the protein fraction in SBM was mostly B2 and FM contained high proportions of B2 and B3 fractions. The proportion of B3 fraction, slowly degradable protein, in CP was the highest in fish meal (23.34%), followed by CMLP (7.68%) and SBM (1.46%). CMLP was degraded up to 51.40% at 18 h of incubation with Streptomyces protease, which was low compared to FM (55.23%) and SBM (83.01%). This may be due to the insoluble portion of CMLP protein being hardly degradable by the protease. The in vitro fermentation by strained ruminal fluid showed that the amount of soluble fraction was larger in CMLP (40.6%) than in SBM (17.8%). However, because the degradation rate constant of the potentially degradable fraction of CMLP (2.0%/h) was lower than that of SBM (5.8%/h), the effective ruminal protein degradability of CMLP (46.95%) was slightly lower than SBM (53.77%). Unavailable fraction in the rumen was higher in CMLP (34.0%) compared to SBM (8.8%). In vitro CP degradability of CMLP by pepsin was 80.37%, which was lower than SBM (94.42%) and FM (89.04%). The evaluation of protein degradability using different approaches indicated that soluble protein in CMLP may supply a large amount of ammonia in the rumen while insoluble protein can be by-passed from microbial attacks due to its low degradability. The results from this study suggest that CMLP can be used as a protein supplement to ruminants for supplying both non-protein nitrogen to rumen microbes and rumen undegradable protein to the host animal.

• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.358-365
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• 2015

ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine molecules connected between the epsilon amino and alpha carboxyl groups. This polymer is currently used as a natural preservative in food. Insufficient biomass is a major problem in ε-PL fermentation. Here, to improve cell growth and ε-PL productivity, various nitrogen-rich nutrients were supplemented into flask cultures after 16 h cultivation, marking the onset of ε-PL biosynthesis. Yeast extract, soybean powder, corn powder, and beef extract significantly improved cell growth. In terms of ε-PL productivity, yeast extract at 0.5% (w/v) gave the maximum yield (2.24 g/l), 115.4% higher than the control (1.04 g/l), followed by soybean powder (1.86 g/l) at 1% (w/v) and corn powder (1.72 g/l) at 1% (w/v). However, supplementation with beef extract inhibited ε-PL production. The optimal time for supplementation for all nutrients examined was at 16 h cultivation. The kinetics of yeast-extract-supplemented cultures showed enhanced cell growth and production duration. Thus, the most commonly used two-stage pH control fed-batch fermentation method was modified by omitting the pH 5.0-controlled period, and coupling the procedure with nutrient feeding in the pH 3.9-controlled phase. Using this process, by continuously feeding 0.5 g/h of yeast extract, soybean powder, or corn powder into cultures in a 30 L fermenter, the final ε-PL titer reached 28.2 g/l, 23.7 g/l, and 21.4 g/l, respectively, 91.8%, 61.2%, and 45.6% higher than that of the control (14.7 g/l). This describes a promising option for the mass production of ε-PL.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.4
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• pp.561-566
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• 2008

Two experiments were conducted to evaluate nutritive value of cell mass from lysine production (CMLP) as a protein supplement for ruminants. In each experiment, animals were fed a diet containing 40% of forages and 60% of concentrates, mainly composed of rice straw and ground corn, respectively, to meet the maintenance requirements, and the diets were formulated to supply equal amounts of energy and nitrogen among treatments. In order to investigate the effect of CMLP on ruminal fermentation (Experiment 1), three Korean native goats weighing $26.1{\pm}1.4kg$ were allotted into individual cages with a $3{\times}3$ Latin square design. Each animal was fed one of three protein sources (CMLP, soybean meal (SBM), and urea). Rumen pH, bacterial and fungal counts, volatile fatty acid concentrations and acetate to propionate ratio were not significantly different among treatments. Concentration of propionate, however, was higher in SBM treatment (14.1 mM) than in CMLP (8.7 mM) or urea (9.3 mM) treatments. There was significantly more branch-chain volatile fatty acid production in CMLP (1.9 mM) and SBM (1.8 mM) treatments than in urea (1.3 mM) treatment. The number of protozoa was the highest in urea treatment, followed by CMLP and SBM treatment with significant differences. A metabolic trial (Experiment 2) was conducted to measure in vivo nutrient digestibility and nitrogen retention in Korean native goats fed CMLP and SBM. Two heavy ($35.0{\pm}1.2kg$) and two light ($25.0{\pm}0.9kg$) Korean native goats, caged individually, were used in this experiment. A heavy and a light animal were paired and supplemented with either CMLP or SBM. The animals fed CMLP showed a trend of lower total tract digestibility in all the nutrients measured; however, there was no statistical significance except for digestibility of ether extract. Nitrogen digestibility of CMLP was estimated to be about 7% units lower than that of SBM. There was a tendency for lower nitrogen retention in CMLP treatment (35.9%) compared to SBM treatment (42.3%). In summary, CMLP can be a good protein source for ruminant animals from nutritional and economic perspectives and may replace some, if not all, of SBM in a diet without losing nitrogen utilization efficiency. Further research is warranted for investigating the effect of CMLP fed with easily fermentable forage and the effective level of CMLP for replacing SBM.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5789-5795
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• 2013

Background: The high mobility group box 1 (HMGB1) protein is a widespread nuclear protein present in most cell types. It typically locates in the nucleus and functions as a nuclear cofactor in transcription regulation. However, HMGB1 can also localize in the cytoplasm and be released into extracellular matrix, where it plays critical roles in carcinogenesis and inflammation. However, it remains elusive whether HMGB1 is relocated to cytoplasm in clear cell renal cell carcinoma (ccRCC). Methods: Nuclear and cytoplasmic proteins were extracted by different protocols from 20 ccRCC samples and corresponding adjacent renal tissues. Western blotting and immunohistochemistry were used to identify the expression of HMGB1 in ccRCC. To elucidate the potential mechanism of HMGB1 cytoplasmic translocation, HMGB1 proteins were enriched by immunoprecipitation and analyzed by mass spectrometry (MS). Results: The HMGB1 protein was overexpressed and partially localized in cytoplasm in ccRCC samples (12/20, 60%, p<0.05). Immunohistochemistry results indicated that ccRCC of high nuclear grade possess more HMGB1 relocation than those with low grade (p<0.05). Methylation of HMGB1 at lysine 112 in ccRCC was detected by MS. Bioinformatics analysis showed that post-translational modification might affect the binding ability to DNA and mediate its translocation. Conclusion: Relocation of HMGB1 to cytoplasm was confirmed in ccRCC. Methylation of HMGB1 at lysine 112 might the redistribution of this cofactor protein.