• Applied Biological Chemistry
• /
• v.14 no.2
• /
• pp.137-148
• /
• 1971

Studies were carried out to investigate the main fermentation microorganisms and their flora changes during Korean native soy-sauce fermentation. Korean native Maeju loaves collected from 5 Do's were separated into surface and inner parts. Four different soy-sauces-the surface part Maeju fermented soy-sauce, the inner part, the surface and inner part combined Maeju fermented soy-sauce, and the semi-Japanese type soy-sauce were fermented and the changes of fermentation microorganism flora and the various chemical components during the period of their fermentations were studied. Besides, 14 home-made soy-sauces collected from 14 different places all over Korea were examined in comparison with the laboratory soy-sauces and to determine the characteristics of Korean native soy-sauce. The results were as follows: 1. The main microorganisms in Korean native soy-sauce fermentation were determined as; Aerobic bacteria: Bacillus subtilis, Bacillus pumilus Lactic acid bacteria: Pediococcus halophilus, Leuconostoc mesenteroides Yeasts: Torulopsis datila, Saccharomyces rouxii 2. Microflora changes during Korean native soy-sauce fermentation were as follows; Aerobic bacteria increased until the 2nd week of fermentation and then gradually decreased. The lactic acid bacteria increased until the 3rd week, after which decreased. When the lactic acid fermentation lowered the pH value to below the 5.4, yeasts were able to grow and participate the fermentation. As the production of organic acids amounted, to a certain height, the growth of all microorganisms lead to the period of decline or death at about the 2nd month of fermentation. After boiling of soy-sauce most microorganisms except a few of Bacillus sp. disappeared. Occosionally yeasts and lactic acid bacteria survived depending upon the composition of soy-sauce. 3. Changes of general chemical components influencing the microflora were investigated for the period of Korean native soy-sauce fermentation. Tetal acidity, salt concentration and total nitrogen were increasing steadily over the entire period of fermentation. pH values were dropping to a certain degree of about 4.5. Salt concentration and pH value seemed to be the important factors influencing the microflora. 4. The microflora were influenced by chemical components of soy-sauce. Aerobic bacteria were able to survive in all soy-sauce as they made spores. Growth of lactic acid bacteria was inhited at 23-26% of salt concentration and pH 4.8. Soy-sauce yeasts started to grow only at pH below 5.4 and seemed to be inhibited at around 26% of salt concentration under pH 4.5-4.7. 5. The open kettle boiling of soy-sauce, the characteristic process of Korean native soy-sauce manufacturing, was effective to sterilize microorganisms, increase the salt concentration, and coagulate proteins. 6. The average viable cell counts of microorganism found in collected samples of home-made Korean native soy-sauces were; Aerobic bacteria: $53{\times}10^2\;cell/ml$ Lactic acid bacteria: 34 cell/ml Yeasts: 14 cell/ml The average values of chemical compositions of samples of home-made Korean native soy-sauce were; Salt concentration: 28.9% pH value: 4.79 Total acidity(lactic acid): 0.91g/100ml Total nitrogen: 1.09g/100ml

• The Korean Journal of Food And Nutrition
• /
• v.26 no.2
• /
• pp.249-257
• /
• 2013

This study is being performed to confirm the container effects during the fermentation processes of kimchi. Kimchi fermentation was prepared in the laboratory with four different types of containers; namely, a traditional Onggi vessel (Korean traditional clay pot, TOV), plastic airtight covered Onggi vessel (PAOV), plastic covered vessel (PCV) and plastic airtight covered vessel (PACV). The kimchi fermentation in the different containers was followed by taking samples at 48 hour intervals for 10 days. In all fermentation containers, the pH changes of kimchi were decreased with fermentation days, while salt content was the same for all types of containers. The number of lactic acid bacteria in kimchi were $1.09{\times}10^8$ $CFU/m{\ell}$ at first. But the TOV, PAOV, PCV, and PACV after fermentation for 10 days were $1.42{\times}10^{10}$, $9.13{\times}10^9$, $4.93{\times}10^9$ and $7.46{\times}10^9$ $CFU/m{\ell}$, respectively. The kimchi fermented in the TOV with the most dominant bacterial species were the following 5 strains: Bacillus subtilis, B. licheniformis, B. safensis, Lactobacillus brevis and B. pumilus. The use of different types of containers therefore influenced the number of L. brevis and the four Bacillus species. in kimchi, and may influence the characteristics of the fermented kimchi products. The TOV offered the greatest L. brevis numbers and suggested that it could be the best suited for preparing traditional kimchi fermentation.

• Asian-Australasian Journal of Animal Sciences
• /
• v.19 no.8
• /
• pp.1164-1173
• /
• 2006

Cellulase from Aspergillus niger, (${\alpha}$-amylase from Bacillus sp. and protease from Bacillus globigii were used as enzyme sources in this study to examine how their respective substrates protect them in two kinds of simulated gastrointestinal tract digesting processes. Avian total digest tract simulation test showed that filter paper, Avicel and cellulose resulted in 7.7, 6.4 and 7.4 times more activity than of unprotected cellulose, respectively. Protease with addition of casein, gelatin or soybean protein showed no significant protection response. Starch protected amylase to be 2.5 times activity of the unprotected one. Monogastric animal total tract digestion simulation test showed that filter paper, Avicel and cellulose resulted in 5.9, 9.0 and 8.8 times activity of unprotected cellulase, respectively. Casein, gelatin and soybean protein resulted in 1.2, 1.3 and 2.0 times activity of unprotected protease, respectively. Starch did not protect amylase activity in monogastric animal total tract simulation. Protection of mixed enzymes by substrates in two animal total tract simulation tests showed that filter paper in combination with soybean protein resulted in 1.5 times activity of unprotected cellulose, but all substrates tested showed no significant protection effect to protease. Soybean protein and starch added at the same time protected the amylase activity to be two times of the unprotected one. Test of non-purified substrate protection in two animal total digest tract simulation showed that cellulase activity increased as BSA (bovine serum albumin) concentration increased, with the highest activity to be 1.3 times of unprotected enzyme. However, BSA showed no significant protection effect to protease. Amylase activity increased to 1.5 times as BSA added more than 1.5% (w/v). Cellulase activity increased to 1.5 times as soybean hull was added higher than 1.5%. Amylase had a significant protection response only when soybean hull added up to 2%. Protease activity was not protected by soybean hull to any significant extent.

• Korean Journal of Food Science and Technology
• /
• v.34 no.4
• /
• pp.666-672
• /
• 2002

Changes in chemical characteristic, microflora, and sensory evaluation of fish sauce extracted at an interval of one week from fermented solution were investigated. pH was reduced from 6.0 to 4.5, and trimethylamine oxide from 132.5 to 87.2 mg/100g during fermenting periods. Trimethylamine increased from 5.6 to 50.2 mg/100g, and volatile basic nitrogen from 48.3 to 232.5 mg/100g. Bacterial flora isolated from the fish sauce were 70% Lactobacillus sp. and 13% Bacillus sp. Among the free amino acids, alanine, glutamic acid, valine, and methionine contents constitute 40% of the total free amino acids. Major non-volatile organic acid of the fish sauce was lactic acid (76%). Sensory evaluation results of the fish sauce were higher than the traditional soybean sauce after 28 days of fermentation.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.4
• /
• pp.730-738
• /
• 2016

est19 is a gene from Bacillus sp. K91 that encodes a new esterase. A comparison of the amino acid sequence showed that Est19 has typical Ser-Gly-Asn-His (SGNH) family motifs and could be grouped into the SGNH hydrolase family. The Est19 protein was functionally cloned, and expressed and purified from Escherichia coli BL21(DE3). The enzyme activity was optimal at 60℃ and pH 9.0, and displayed esterase activity towards esters with short-chain acyl esters (C₂-C₆). A structural model of Est19 was constructed using phospholipase A1 from Streptomyces albidoflavus NA297 as a template. The structure showed an α/β-hydrolase fold and indicated the presence of the typical catalytic triad Ser49-Asp227-His230, which were further investigated by site-directed mutagenesis. To the best of our knowledge, Est19 is a new member of the SGNH hydrolase family identified from thermophiles, which may be applicable in the industrial production of semisynthetic β-lactam antibiotics after modification.

• Korean Journal of Soil Science and Fertilizer
• /
• v.37 no.2
• /
• pp.116-123
• /
• 2004

This study was conducted to investigate the changes in physicochemical and microbiological properties during fermenting process of organic fertilizer which was made from the mixture of organic materials such as sesame oil cake, fish meal, blood meal, rice bran, ground bone meal, and natural minerals such as illite, crusted oyster shell and loess. They were mixed and fermented for 70 days. The sesame oil cake and rice bran, major ingredients for organic fertilizers, consisted of 7.6 and 2.6% total nitrogen, 3.6 and 4.6% $P_2O_5$, 1.4 and 2.2% $K_2O$, respectively. The ground bone meal included 29.2% $P_2O_5$ and illite included 3.8% $K_2O$. Temperature of organic fertilizer during the fermentation rapidly increased over $50^{\circ}C$ within 2 days after mixing and stabilized similar to outdoor temperature after 40 days. Moisture content decreased from 36.3 to 16.0% after 1 month. C/N ratio of organic fertilizer slightly increased until 30 days and thereafter, it slowly decreased, It resulted from the faster decrease of total nitrogen concentration compared with organic matter. Concentration of $NH_4-N$ in organic fertilizer rapidly increased from 1,504 to $5,530mg\;kg^{-1}$, the highest concentration after 10 days. Meantime, $NO_3-N$ concentration was low and constant about $150mg\;kg^{-1}$ over the whole fermenting period. This result seemed to be due to the high pH. The organic ferfilizer fermented for 70 days was composed of 2.7% N, 2.8% $P_2O_5$, 1.8% $K_2O$, and 35.9% organic matter. Total populations of aerobic bacteria, Bacillus sp. and actinomycetes, after fermenting process, were $12.5{\times}10^{10}$, $45.5{\times}10^{5}$ and $13.6{\times}10^{5}cfu\;g^{-1}$ respectively. Pseudomonas sp. was $71.9{\times}10^{7}cfu\;g^{-1}$ at first, but it rapidly decreased according to the rise of temperature. Yeasts played an important role in the early stage of fermentation and molds did in the late stage.

• Microbiology and Biotechnology Letters
• /
• v.39 no.4
• /
• pp.330-336
• /
• 2011

Bacterial strains exhibiting fibrinolytic activity were screened from traditional Korean soybean sauce. The Fibrinolytic activities of the various isolated microorganism were further examined and the superior strain YJ11-21 was selected for further analyses. Gene sequence analysis of 16S rDNA of the YJ11-21 strain revealed Bacillus licheniformis. Optimal culture conditions were investigated in order to maximize the production of the fibrinolytic enzyme by YJ11-21. Amongst the carbon sources tested, glucose was the most effective for enzyme production and amongst the nitrogen sources tested, yeast extract was seen to be the most effective. A one percent addition of NaCl to the medium resulted in the highest fibrinolytic activity. Interestingly, a 10% addition of NaCl resulted in a high activity together with a high cell growth rate. Therefore, YJ11-21 is speculated of being a halotolerant. The optimum pH and temperature for enzyme production were a pH of 9.0 and $30^{\circ}C$, respectively.

• Journal of Microbiology
• /
• v.40 no.1
• /
• pp.33-37
• /
• 2002

An enzymatic reaction for the production of D-alanine from D-aspartic acid and pyruvate as substrates by a thermostable D-amino acid aminotransferase (D-AAT) was investigated at various conditions In the temperature range of 40-70$\^{C}$ and pH range of 6.0-9.5. The D-AAT was produced with recombinant E. coli BL21, which hosted the chimeric plasmid pTLK2 harboring the D-AAT from the novel thermophilic Bacillus sp. LK-2. The enzyme reaction was shown to follow the Ping Pong Bi Bi mechanism. The K$\_$m/ values for D-aspartic acid and pyruvate were 4.38 mar and 0.72 mM, respectively. It was observed that competitive inhibition by D-alanine, the product of this reaction, was evident with the inhibition constant K$\_$i/ value of 0.1 mM. A unique feature of this reaction scheme is that the decorboxylation of oxaloacetic acid, one of the products, spontaneously produces pyruvate. Therefore, only a catalytic amount of pyruvate is necessary for the enzyme conversion reaction to proceed. A typical time-course kinetic study skewed that D-alanine up to 88 mM could be produced from 100 mM of D-aspartic acid with a molar yield of 1.0.

• Journal of Life Science
• /
• v.27 no.2
• /
• pp.202-210
• /
• 2017

As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

• Korean Journal of Food Science and Technology
• /
• v.4 no.4
• /
• pp.276-284
• /
• 1972

A strain of Bacillus sp. which has powerful enzymatic activity and sets free a little amount of ammonia nitrogen and large amount of amino nitrogen was separated from Meju, Denzang, Chungkukjang, and paddy straw to make the soy-sauce rapidly and conveniently in the various mixing ratio of steamed soy-bean and wheat parched or steamed. Total nitrogen and amino nitrogen were increased during the maturing. The acidity of soy-sauce was increased to $1.15{\sim}1.98%$ than ordinary soy-sauce. pH were ranged in $4.6{\sim}6.0%$. The fermenting time takes 30 days while it takes at least 4 monthes in ordinary method. The best results were obtained with the mixed ratio of 1 : 1 or 1 : 2 (wheat : soy-bean).