• Applied Biological Chemistry
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• v.42 no.2
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• pp.83-90
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• 1999

Four strains of lactic acid bacteria which had cholesterol lowering activities were selected from foreign fermented milk. The strains were identified as Lactobacillus(L.) rhamnosus 2084, L. casei 0781, Lactococcus (Lacto.) lactis spp. 204, and Enterococcus(E.) faecium 402. We observed that the L. rhamnosus 2084 was the most tolerant against pH 1.5, L. casei 0781 against pH 2.0, but not significantly different in the tolerance against pH 3.0. The L. rhamnosus 2084 was the most tolerant against bile acid and prominent in the degree of lowering cholesterol level. All four strains were used as starters in producing yogurt, and then investigated physico-chemical characteristics, such as pH, titratable acidity, and viable cell counts of yogurt base. L. casei 0781, L. rhamnosus 2084, Lacto. lactis 204, and E. faecium 402 were incubated for 6 hours at $40^{\circ}C$, 4 hours at $40^{\circ}C$, 6 hours at $37^{\circ}C$, and 12 hours at $37^{\circ}C$ and $40^{\circ}C$, respectively, for the optimum conditions of fermented milk.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1471-1480
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• 2010

An extremely thermostable xylanase gene, xynB, from the hyperthermophilic bacterium Thermotoga maritima MSB8 was successful expressed in Kluyveromyces lactis. The response surface methodology (RSM) was also applied to optimize the medium components for the production of XynB secreted by the recombinant K. lactis. The secretion level (102 mg/l) and enzyme activity (49 U/ml) of XynB in the optimized medium (yeast extract, lactose, and urea; YLU) were much higher than those (56 mg/l, 16 U/ml) in the original medium (yeast extract, lactose, and peptone; YLP). The secretory efficiency of mature XynB was also improved when using the YLU medium. When the mRNA levels of 13 characterized secretion-related genes in the K. lactis cultured in YLP and YLU were detected using a semiquantitative RT-PCR method, the unfolded protein response (UPR)-related genes, including ero1, hac1, and kar2, were found to be up-regulated in the K. lactis cultured in YLU. Therefore, the nutrient ingredients, especially the nitrogen source, were shown to have a significant influence on the XynB secretory efficiency of the host K. lactis.

• Microbiology and Biotechnology Letters
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• v.49 no.1
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• pp.75-87
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• 2021

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp₄₅ signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SP_Usp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC₅₀ values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

• Applied Biological Chemistry
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• v.46 no.4
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• pp.367-375
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• 2003

The puree of Rubus coreanus was fermented using lactic acid bacteria and its biological activities were examined. Lactobacillus acidophilus KCCM 32820, L. casei KCCM 12452, Lactococcus lactis subsp. lactis KCCM 40104, and Streptococcus thermophilus KCCM 40430 were used as a single or mixed starter for the lactic acid fermentation, and their cultures at the late logarithmic growth phase were inoculated to final concentration of 2% (v/v). L. casei fermented the puree of Rubus coreanus best when used as a single starter, and the culture of L. casei and L. lactis with the inoculation ratio of one to one showed the highest fermentation activity when used as a mixed starter. However, the fermented broth of the puree of Rubus coreanus using L. acidophilus and S. thermophilus showed the best results in the sensory evaluation. The optimal lactic acid fermentation conditions were as follows; the concentration of oligosaccharide added was 1% (w/v), pH of puree and fermentation temperature were 4.0 and $37^{\circ}C$, respectively, and fermentation time was $72{\sim}96$ hours. Glucose and fructose were major free sugars, and the content of lactic acid was 698.2 mg/100 g in the fermented broth. The fermented broth of the puree of Rubus coreanus showed the electron donating ability and nitrite scavenging ability with the value of 69% and 38.3% at pH 1.2, respectively. SOD-like activity and inhibitory activity on xanthine oxidase were also found in the fermented broth with the value of 60.3% and 41.8%, respectively. When the antimicrobial activities of the fermented broth were examined, it showed the highest growth inhibitory activity against Escherichia coli O-157:H7, and also contained antimicrobial activities against Salmonella typhimurium, Bacillus cereus, and Staphylococcus aureus.

• Journal of Applied Biological Chemistry
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• v.62 no.3
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• pp.257-264
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• 2019

Rice bran is considered a natural source of antioxidants. In this study, rice bran was fermented with lactic acid bacteria to increase its antioxidant activity. Four strains isolated from fermented food, Lactobacillus plantarum MJM60383, Lactococcus lactis subsp. lactis MJM60392, Lactobacillus fermentum MJM60393, and Lactobacillus paracasei MJM60396, were confirmed as safe through stability tests such as safety assessment for biogenic amine production, hemolytic activity, and mucin degradation, and showed high reducing capacity. The antioxidant activity of rice bran fermentation altered by these strains was evaluated using several methods including measurement of $Fe^{2+}$ chelating activity and scavenging activity by 1,1-diphenyl-2-picryl-hydrazil (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and nitric oxide assays. In this study, the total phenolic content and ${\gamma}$-oryzanol were evaluated by high-performance liquid chromatography. Compared to non-fermented rice bran and a commercial product, rice bran fermented with Lactococcus lactis subsp. lactis MJM60392 showed the highest phenolic content (844.13 mg GAE/g). Moreover, the content of ferulic acids, ${\rho}$-coumaric acid, and ${\gamma}$-oryzanol in rice bran increased after fermentation with L. lactis subsp. lactis MJM60392 and L. fermentum MJM60393 compared to other samples. Indeed, the DPPH radical scavenging activity and NO scavenging activity were also found to be high in these fermented rice brans. These results indicated that fermentation with lactic acid bacteria increases the active compound levels and the potent antioxidant activities of rice bran.

• Korean Journal of Food Science and Technology
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• v.46 no.2
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• pp.165-172
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• 2014

Mixed fermentation of cow milk was performed by sequential co-cultures with Bacillus subtilis and Lactococcus lactis. After a first fermentation step with B. subtilis for 6 h, the number of viable cells increased to $2.5{\times}10^8$ CFU/mL. The second fermentation step with L. lactis resulted in increased viable cells $1.09{\times}10^{10}$ CFU/mL for 3 days and increased acidity. However, the number of viable B. subtilis cells was decreased greatly to $5{\times}10^1$ CFU/mL following fermentation with L. lactis. Milk proteins were markedly hydrolyzed by the first fermentation after 2 h, and the second fermentation induced curd formation in milk. However, after 4 h, the first fermentation resulted in higher whey separation and 80 mg% tyrosine content. Gamma-aminobutyric acid (GABA) production was dependent upon the degree of protein hydrolysis by first fermentation. Second fermentation resulted in 0.14% GABA. The milk fermented by B. subtilis indicated the rough surface of yogurt depended upon the degree of protein hydrolysis. In conclusion, set-type yogurt was efficiently produced by co-culturing of milk, and fortifying with peptides, GABA, and probiotics.

• Korean Journal of Food Science and Technology
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• v.14 no.1
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• pp.57-62
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• 1982

The possibility of developing new kinds of lactic acid beverage from a malt syrup was studied. The optimum sugar concentration of malt syrup for the cultivation of lactic acid bacteria was $10^{\circ}Bx$. The acidity of the fermented malt syrup was improved by the supplement of yeast extract(0.5%) or sodium citrate(0.08%). Though the activity of Lactobacillus lactis in malt syrup was superior to other strains, sensory test indicated that the mixed culture of Lactobacillus lactis and Streptococcus diacetilactis was better because of masking malt flavour. The changes in acidity and viable cells of malt syrup during the lactic fermentation were not so good as skim milk medium, but malt syrup medium containing milk(50 : 50) was nearly similar to skim milk medium. In the sensory scores among samples, no significant differences(P<0.05) were noted between fermented milk and fermented malt syrup containing milk, but fermented malt syrup showed a poor quality. However fermented malt syrup was not inferior to marketing lactic fermented fruit juices with regards to the lactic acid fermented beverage type.

• Food Engineering Progress
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• v.13 no.1
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• pp.64-69
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• 2009

From the 25 fresh fruits and vegetable products, 1,250 isolates grown on MRS agar media were screened for the inhibitory activity on the growth of Escherichia coli 0157:H7, Listeria monocytogenes, and Bacillus cereus as well as Lactobacillus plantarum, L. casei, and Lactococcus lactis subsp. lactis. Among them, 607 isolates (49% of total isolates) from 23 different foods produced growth inhibitory activity on the E. coli 0157:H7, L. monocytogenes, or B. cereus. When these isolates were screened for the inhibitory activity on the growth of L. plantarum, L. casei, and Lactococcus lactis subsp., 24 isolates (3% of total isolates) from 7 food samples showed bacteriocin-like activity. These isolates had typical physiological characteristics of lactic acid bacteria, which indicated these isolates were strains of lactic acid bacteria. The inhibitor from 3 out of 24 revealed as nicin. From the RAPD-PCR profiles, 24 strains was classified and it was also indicated that most of the strains isolated from same produce showed similar phylogenetic profile.

• The Korean Journal of Food And Nutrition
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• v.14 no.2
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• pp.126-131
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• 2001

Microbiological, pH and sensory evaluations of pork loins treated with lactic acid cultures (Lactococcus lactis subsp. lactis ATCC 11454 grown in 10% milk solids) during storage at 4$^{\circ}C$ were assessed. Pork loins were immersed in solutions containing 3.96 log units, 4.10 log units, or 4.23 log units of individual lactic acid cultures for 2 min. Pork loins treated with lactic acid cultures had a inhibitory effect (P<0.05) on the growth of aerobic palate counts during storage of 3 days at 4$^{\circ}C$. Pork loins treated with lactic acid cultures during storage of 3 days at 4$^{\circ}C$ significantly (P<0.05) decreased pH values compared to those of controls. Sensory evaluations for odor and appearance of pork loins treated with lactic acid cultures during storage of 9 days at 4$^{\circ}C$ were a "liked less" to "typical" category compared to those of controls.

• Applied Biological Chemistry
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• v.23 no.4
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• pp.218-221
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• 1980

From S. lactis $KB_{21}$, stabilized strain by intergration of the lactose plasmid into the host chromosome, several lactose constitutive mutants were obtained by UV irradiation and spontaneous mutation. The rapid growth of the mutants in the medium containing lactobionate confirmed their constitutive nature. The mutants synthesized $phospho-{\beta}-galactosidase$ with an activity of 1.7 to 3.4 times that of the parent.