• Korean Journal of Microbiology
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• v.54 no.4
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• pp.369-378
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• 2018

This study was conducted to evaluate effects of dietary microbial probiotics on the growth and disease resistance of olive flounder (Paralichthys olivaceus) in a recirculating aquaculture system (RAS), and the effects of the probiotic bioaugmentation on the microbial community structure and water quality. For the analysis, 80 juvenile fish (average weight, $25.7{\pm}7.6g$; average length, $15.2{\pm}1.7cm$) were fed a basal diet containing a commercial microbial product CES-AQ1 (CES; $1{\times}10^9\;CFU/kg$ diet) in an RAS for 8 weeks. Weight gain, the specific growth rate, feed efficiency, and protein efficiency ratio of the fish fed the CES diet in the RAS were 1.5~2.5 times higher than those of fish fed the basal diet alone, or the basal diet containing oxytetracycline (OTC), yeast plus bacterium, or Bacillus subtilis in a still water system. There was no significant difference in the pathogen challenge test between fish fed the OTC diet and fish fed the CES diet in the RAS, suggesting the CES-AQ1 probiotic used in the RAS as a potential replacement for antibiotics. The RAS biofilter maintained the highest microbial diversity and appeared to harbor microbial communities with ammonium oxidation, denitrification, and fish pathogen suppression functions. Ammonia, which is hazardous to fish, was significantly decreased to < 0.5 mg/L in 19 days, indicating the effectiveness of probiotic supplementation to maintain good water quality in RAS. These results suggest that the intestinal microbial communities of fish are stabilized by a probiotic-containing diet (CES) and that bioaugmentation with probiotics may be an eco-friendly and economical supplement for aquaculture of olive flounder, promoting both good water quality and fish health in an RAS.

• Journal of Food Hygiene and Safety
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• v.37 no.2
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• pp.87-96
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• 2022

Recently, the purchase of fresh-cut produce and meal kits has increased. Ready-to-eat (RTE) fresh-cut products have potentially hazard of cross-contamination of various microorganisms in the processes of peeling, slicing, dicing, and shredding. There are frequent cases of protozoa food poisoning, such as Cyclospora and Cryptosporidium, caused by fresh-cut products. The objective of the study is to investigate the microbiological qualities of various types of RTE fresh-cut products in the domestic on/offline markets. RTE fresh-cut fruits cup (n=100), fresh-cut vegetables (n=50), and vegetables in meal kits (Vietnamese spring rolls and white radish rolls kits, n=50) were seasonally analyzed. The contamination levels of hygienic indicator organisms, yeast and mold (YM), and foodborne pathogens (Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7) were monitored. Overall, the lowest microbiological qualities of meal kits vegetables were observed, followed by RTE fresh-cut fruits cup and fresh-cut vegetables. Contamination levels of total aerobic bacteria, coliforms, and YM in meal kits vegetables were 5.91, 3.90, and 4.71 logs CFU/g, respectively. From the qualitative analysis, 6 out of 200 RTE fresh-cut products (3%) returned positive result for S. aureus. From the quantitative analysis, the contamination levels of S. aureus in purple cabbage from a meal-kit and fresh-cut pineapple were below the acceptable limit (100 CFU/g). Staphylococcus enterotoxin seg and sei genes were detected in RTE fresh-cut celery and red cabbage from meal-kits, respectively. S. aureus contamination must be carefully controlled during the manufacturing processes of RTE fresh-cut products. Neither Cyclospora cayetanensis nor Cryptosporidium parvum was detected in the samples of RTE fresh-cut products and vegetables from meal-kits from the Korean retail markets.

• Research in Plant Disease
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• v.28 no.2
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• pp.69-81
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• 2022

This study was performed to screen the efficacy of antagonistic bacterial isolates from various sources against the bacterial fruit blotch (BFB) causing pathogen (Acidovorax citrulli) in cucurbit crops. In addition, plant growth promoting traits of these antagonistic bacterial isolates were characterized. Two thousand seven hundred ninety-four microorganisms were isolated from the collected samples. Molecular identification revealed two A. citrulli out of 2,794 isolates. In vitro antagonistic results showed that, among the 28 antagonistic bacterial isolates, 24 and 14 bacterial isolates exhibited antagonism against HPP-3-3B and HPP-9-4B, respectively. Antagonistic and growth promotion characterization of the antagonistic bacterial isolates were further studied. Results suggested that, 4 antagonistic bacteria commonly showed both antagonism and growth promotion phenotypes. Moreover, 3 isolates possessed growth promoting activities. Overall results from this study suggests that BFB causing bacterial pathogen (A. citrulli) was suppressed in in vitro antagonism assay by antagonistic bacterial isolates. Furthermore, these antagonistic bacterial isolates possessed growth promotion and antagonistic enzyme production ability. Therefore, data from this study can provide useful basic data for the in vivo experiments which ultimately helps to develop the eco-friendly agricultural materials to control fruit rot disease in cucurbit crops in near future.

• Journal of Life Science
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• v.33 no.1
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• pp.1-7
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• 2023

Intracellular cargo transport is mediated by molecular motor proteins, such as kinesin and cytoplasmic dynein. Kinesins make up a large subfamily of molecular motors. Kinesin-1 is a plus-end-directed molecular motor protein that moves various cargoes, such as organelles, protein complexes, and mRNAs, along a microtubule track. It consists of the kinesin superfamily protein (KIF) 5A, 5B, and 5C (also called kinesin heavy chains) and kinesin light chains (KLCs). Kinesin-1 interacts with many different binding proteins through its carboxyl (C)-terminal region of KIF5s and KLCs, but their binding proteins have not yet been fully identified. In this study, a yeast two-hybrid assay was used to identify the proteins that interact with the KIF5A specific C-terminal region. The assay revealed an interaction between KIF5A and glutamate-rich 4 (ERICH4). ERICH4 bound to the KIF5A specific the C-terminal region but did not interact with the C-terminal region of KIF5B or KIF3A (a motor protein of kinesin-2). In addition, KIF5A did not interact with another isoform, ERICH1. Glutathione S-transferase (GST) pull-downs showed that KIF5A interacts with GST-ERICH4 and GST-ERICH4-amino (N)-terminal but not with GST-ERICH4-C or GST alone. When co-expressed in HEK-293T cells, ERICH4 co-localized with KIF5A and co-immunoprecipitated with KIF5A and KLC but not KIF3B. Together, our findings suggest that ERICH4 is capable of binding to KIF5A and that it may serve as an adaptor protein that links kinesin-1 with cargo.

• Journal of Life Science
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• v.32 no.3
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• pp.189-195
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• 2022

Kinesin-2 comprises two subfamilies of the heterotrimeric or homodimeric motors found in mammalian cells. Heterotrimeric kinesin-2 consists of kinesin superfamily proteins (KIFs) 3A and 3B and kinesin-associated protein 3 (KAP3), which is a molecular motor protein that moves along microtubules. It plays diverse roles in cargo transport, including anterograde trafficking in cilia, and interacts with many different cargoes and proteins, but their binding proteins have not yet been fully identified. In this study, the yeast two-hybrid assay was used to identify the proteins that interact with the cargo-binding domain (CBD) of KIF3A, and an interaction between KIF3A and brain expressed X-linked 2 (Bex2) was found. Bex2 bound to the CBD-containing C-terminal tail region of KIF3A but did not interact with the same region of KIF3B or KIF5A (a motor protein of kinesin-1). KIF3A interacted with another isoform, Bex1, but did not interact with Bex3. In addition, glutathione S-transferase (GST) pull-downs showed that KIF3A specifically interacts with GST-Bex1 and GST-Bex2 but not with GST alone. When co-expressed in HEK-293T cells, Bex2 co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. In combination, these results suggest that Bex2 is capable of binding to heterotrimeric kinesin-2 and may serve as an adaptor protein that links heterotrimeric kinesin-2 with cargo.

• Journal of Life Science
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• v.33 no.7
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• pp.531-537
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• 2023

Intracellular and axonal transport is mediated by microtubule-dependent motor proteins, such as kinesins and cytoplasmic dynein. Kinesin moves along the microtubule to the positive end of the microtubule, while dynein moves to the negative end of the microtubule. Kinesin-1 was first identified as a kinesin superfamily protein (KIF) that functions in the intracellular transport of various cargoes, including organelles, neurotransmitter receptors, and mRNA-protein complexes, through interactions between the carboxyl (C)-terminal domain and the cargo. It interacts with other cargoes, but the adapter/scaffold proteins that mediate between kinesin-1 and the cargo have yet to be fully identified. In this study, a yeast two-hybrid screen was used to identify adapter proteins that interact with the C-terminal region of KIF5A. We found an association between the C-terminal region of KIF5A and the cyclin-dependent kinase 2-associated protein 1 (CDK2AP1), originally identified in malignant hamster oral keratinocytes. CDK2AP1 bound to the C-terminal region of KIF5A and did not interact with KIF3A (the motor of kinesin-2), KIF5B, KIF5C, and kinesin light chain 1 (KLC1). The C-terminal region of CDK2AP1 is essential for its interaction with KIF5A. When co-expressed in HEK-293T cells, CDK2AP1 and kinesin-1 co-immunoprecipitated and co-localized in the cells. These results suggest that the KIF5A-CDK2AP1 interaction serves as an adapter protein connecting kinesin-1 and the cargo when kinesin-1 transports cargo in cells.

• Journal of Life Science
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• v.33 no.11
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• pp.868-875
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• 2023

Kinesin-1 is a motor protein identified as the first member of the kinesin superfamily (KIF), which plays a role in intracellular cargo transport by acting as microtubule-dependent motor proteins within cells. Kinesin-1 consists of two heavy chains (KHCs, also known as KIF5s) and two light chains (KLCs). The 93 amino acids in the carboxyl (C)-terminal tail region of KIF5A are not homologous to the C-terminal tail region of KIF5B or the C-terminal tail region of KIF5C. In this study, we used a yeast two-hybrid screen to identify the binding proteins that interacted with the C-terminal region of KIF5A. We found an association between KIF5A and CUE domain containing 2 (CUEDC2), which is proposed to function as an adaptor protein involved in ubiquitination pathways and protein trafficking. CUEDC2 bound to the C-terminal region of KIF5A and did not interact with KIF5B (the motor of kinesin-1), KIF3A (the motor of kinesin-2), or kinesin light chain 1 (KLC1). KIF5A specifically bound to the C-terminal region of CUEDC2. Furthermore, KIF5A did not interact with another isoform: CUEDC1. In addition, glutathione S-transferase (GST) pull-downs showed that KIF5A directly bound GST-CUEDC2 but did not interact with GST-CUEDC1 and GST alone. When myc-KIF5A and EGFP-CUEDC2 were co-expressed in HEK-293T cells, CUEDC2 co-immunoprecipitated with kinesin-1, and myc-KIF5A and FLAG-CUEDC2 colocalized in the cells. These results suggest that in intracellular cargo transport by kinesin-1, CUEDC2 serves as an adaptor protein connecting kinesin-1 and cargo by binding to KIF5A.

• Food Science and Preservation
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• v.30 no.1
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• pp.146-154
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• 2023

To develop a multi-functional ingredient, the bioconversion of katsuobushi protein was optimized using Bacillus subtilis HA and Lactobacillus plantarum KS2020. The Dendropanax morbiferus extract (DME) culture with protease activity (102 unit/mL) was prepared by B. subtilis with 2% glucose and 1% skim milk through one day of alkaline fermentation. Katsuobushi protein was effectively hydrolyzed by the DME culture at 60℃ for 3 hours, resulting in a tyrosine content of 156.85 mg%. Subsequently, a second lactic acid fermentation was carried out with 10% monosodium glutamate (MSG) using L. plantarum KS2020 to produce higher levels of GABA. Following co-cultivation for three days, DME exhibited a pH of 8.3 (0% acidity). After seven days, the viable cell count of L. plantarum increased to 9.33 CFU/mL, but viable Bacillus cells were not detected. Taken together, a multi-functional ingredient with enriched GABA, peptides, probiotics, and umami flavor was developed through lactic acid fermentation using hydrolyzed katsuobushi protein. These results indicate that katsuobushi protein could be used as a byproduct to produce a palatable protein hydrolysate using alkaline-fermented DME culture as a proteolytic enzyme source.

• Food Science and Preservation
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• v.30 no.4
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• pp.703-715
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• 2023

This study was conducted to investigate the fermentation characteristics and anti-obesity effects of acetic acid fermentation products of coffee wine. The live cell counts, soluble solids, pH and total acidity of the acetic acid unfermented coffee wine (AUFCW; day 0, before fermentation) were 6.35 log CFU/mL, 8.10 °Brix, 3.88, and 1.29%, respectively, while the acetic acid fermented coffee wine (AFCW; day 15, after fermentation) were 4.40 log CFU/mL, 8.57 °Brix, 3.07, and 7.45%, respectively. Pancreatic lipase inhibitory activity tended to increase as the acetic acid fermentation period increased. The anti-obesity effects of AFCW on 3T3-L1 cells, which was induced by MDI, were evaluated based on the lipid accumulation rate, leptin expression, and fat production-related gene expression (PPAR-γ and SREBP-1c) at the mRNA level. In the case of AFCW, the lipid accumulation rate and leptin expression were decreased to 69.37% and 50.20% at a concentration of 200 ㎍/mL, respectively, and the expression levels of PPAR-γ and SREBP-1c at the mRNA level were decreased to 79.89% and 48.81%, respectively. These results indicate that anti-obesity effect of acetic acid fermentation products could be increased by acetic acid fermentation of coffee wine.

• Applied Biological Chemistry
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• v.11
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• pp.1-27
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• 1969

More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.