• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.17-23
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• 2009

The gene encoding Thermus thermophilus HJ6 DNA polymerase (Tod) was cloned and sequenced. The open reading frame (ORF) of the Tod gene was composed of 2,505 nucleotides and encoded a protein (843 amino acids) with a predicted molecular weight of 93,795 Da. The deduced amino acid sequence of Tod showed 98% and 86% identities to the Thermus thermophilus HB8 DNA pol and Thermus aquaticus DNA pol, respectively, The Tod gene was expressed under the control of the bacteriophage $\lambda$ promoters PR and PL on the expression vector pJLA503 in Escherichia coli strain BL21 (DE3) codon plus. The expressed enzyme was purified by heat treatment, $HiTrap^{TM}$ Q column, and $HiPrep^{TM}$ Sephacryl S-200 HR 26/60 column chromatographies. The optimal temperature and pH for DNA polymerase activity were found to be $75{\sim}80^{\circ}C$ and 9.0, respectively. The optimal concentrations of $Mg^{2+}$ and $Mn^{2+}$ were 2.5 mM and 1 mM, respectively. The enzyme activity was activated by divalent cations, and was inhibited by monovalent cations. The result of the PCR experiment with Tod DNA polymerase indicates that this enzyme might be useful in DNA amplification and PCR-based applications.

• Journal of Dairy Science and Biotechnology
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• v.33 no.1
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• pp.17-25
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• 2015

The present study was carried out to evaluate the preparation of the fermented milks with rice bran and to prove that the bacteria used are necessary for providing amino acids in this process. The rice bran on fermented milk with Streptococcus thermophilus (ST-body1) and Lactobacillus casei (LC-10). The fermentation limit was set until acidimetry score reaches 1. There are reports of titratable acidity, pH, viable cell count and amounts of organic acids affecting amino acid production about physical and chemical analysis measured using HPLC. Finally, sensory test was surveyed. In this study, the rate of acidification was higher in the fermented milk with rice bran than in the common fermented milk. In case of the number of cells was $1.0{\times}10^8CFU/mL$ in group. The lactic acid and citric acid content in yogurts prepared with rice bran using Streptococcus thermophilus (ST-body1) and Lactobacillus casei (LC-10) was higher than that in the control yogurt. Amino acids derived by rice bran were effected in fermentation for each bacteria's necessary amino acid production, and it made bacteria growth larger. From the physical test of the fermented milk with rice bran, flavor, texture, sweetness, overall taste of the fermented milk of Streptococcus thermophilus (ST-body1) were found to be much better than those of the other groups. The results obtained for the fermented milk prepared with rice bran using Streptococcus thermophilus (ST-body1) are significant.

• Korean Journal of Food Science and Technology
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• v.18 no.6
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• pp.492-496
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• 1986

A compound stimulatory to the growth of S. thermophilus CH-1 was isolated from the cell-free filtrate of L. helveticus YM-1 in milk medium. The stimulant was identified as a peptide with a molecular weight of approximately 5000 and exhibited positive ninhydrin reaction. Some kinds of amino acids confirmed as aspartic acid, alanine, valine, glutamic acid, lysine, proline, leucine were rich in the stimulatory peptide hydrolysate. Among them, glutamic acid was most abundant. Judging from bioautographic results, glutamic acid and phenylalanine were expected to exert an important role for the stimulation.

• Preventive Nutrition and Food Science
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• v.6 no.2
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• pp.96-100
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• 2001

Mixtures prepared from whole milk with added skin milk powder(2.5%, w/v) and Angelica keiskei juice (1.5%, w/v) were fermented with lactic acid bacteria (single and mixed culture of Lactobacillus bulgaricus and Strpetococcus thermophilus) for 24 hours. The fermented mixtures (curd yogurt) were evaluated for acid production (pH and titratable acidity), cell numbers, viscosity, sensory property and keeping quality. Results indicated that the addition of Angelica keiskei stimulated the acid production by lactic acid bacteria. The number of viable cells reached 4.5~7.3$\times$10$^{9}$ CFU/mL for Angelica keiskei-added yogurts, while 3.3~5.1$\times$10$^{9}$ CFU/mL for control yogurts. Viscosity of Angelica keiskei-added yogurts was higher (3,609~3,854 centipoises) than that of control yogurts(3,346~3,700 centipoises). Of the microorganism tested, mixed culture of Lactobacillus bulgaricus and Streptococcus thermophilus was most effective in acid production. The overall sensory score showed that Angelica keiskei yogurt fermented with Streptococcus thermophilus was evaluated as good as control yogurt. When yogurts were stored at 4$^{\circ}C$ for 12 days, pH, titratable acidity and viable cells of lactic acid bacteria were not significantly changed(p<0.05).

• KSBB Journal
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• v.27 no.4
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• pp.257-262
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• 2012

The gene encoding Thermus thermophilus HJ6 laccase (Tt-laccase) was cloned, sequenced, and comprised of 1,389 nucleotides encoding a protein (462 amino acids) with a predicted molecular mass of 51,049 Da. The deduced amino acid sequence of Tt-laccase showed 99.7% and 44.3% identities to the Thermus thermophilus HB27 laccase and Synechococcus sp. RS9917 laccase, respectively. Tt-laccase gene was expressed as a fusion protein with six histidine residues in E. coli Rosetta-gami (DE3) cells, and the recombinant protein was purified to homogeneity. UV-Vis spectrum analysis revealed that the enzyme has copper atoms, a type I Cu(II) and a type III binuclear Cu(II). The optimum pH for the oxidation of guaiacol was 5.0 and the optimum temperature was $90^{\circ}C$ The half-life of heat inactivation was about 120 min at $90^{\circ}C$ The enzyme reaction was inhibited by sodium azide, L-cystein, EDTA, dithiothreitol, tropolone, and kojic acid. The enzyme oxidized various known laccase substrates, its lowest $K_m$ value being for 4-hydroxyindole, highest $k_{cat}$ value for syringaldazine, and highest $k_{cat}/K_m$ for guaiacol.

• Korean journal of food and cookery science
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• v.15 no.1
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• pp.73-80
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• 1999

This study was carried out to investigate the effects of enzyme treatments on the functional properties of soy protein isolate (SPI) and to examine the quality attributes of soy yogurt prepared by different enzyme treatments, sweeteners and starter cultures. Enzyme treatment increased the solubility and emulsifying capacity of soy proteins, but decreased the emulsifying stability; the enzymatic activity of ${\alpha}$-chymotrypsin was higher than that of trypsin. Enzyme treatments decreased the pH of soy yogurts prepared by both culture methods, the culture of L. bulgaricus and S. thermophilus and the culture of L. bulgaricus and K. fragilis, but increased the titratable acidity, total numbers of lactic acid bacteria and yeast. Trypsin was more effective than ${\alpha}$-chymotrypsin in decreasing pH and increasing titratable acidity and total numbers of lactic acid bacteria and yeast. Fructose decreased the pH of soy yogurts more than sucrose in the culture of L. bulgaricus and S. thermophilus, and vice versa in the culture of L. bulgaricus and K. fragilis. Fructooligosaccharides were more effective in the culture of L. bulgaricus and K. fragilis than in the culture of L. bulgaricus and S. thermophilus in increasing the titratable acidity, total count of lactic acid bacteria and yeast. In sensory evaluation, soy yogurts containing trypsin treated SPI, fructose and fructooligosaccharides (75%:25%) were more acceptable than those containing untreated or trypsin treated SPI and fructose. This was because of more smooth and less sour, in which the values of pH, titratable acidity, microbial growth, and viscosity were in the range of commercial yogurts. Soy yogurts fermented by L. bulgaricus and K. fragilis showed more smooth mouthfeel than those fermented by L. bulgaricus and S. thermophilus.

• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.35-45
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• 1989

Streptococcus thermophilus 510 was investigated as n potential source of $\beta$-galactosidase. Optimum cultural conditions for maximum enzyme production were 0.5% loctose as carbon source, initial pH 7.0, 37 $^{\circ}C$, and 18 hours of cultivation. The enzyme was purified to homogeneity by ammonium sulfate fractionation, protamine sulfate precipitation, Sephadex G-200 gel filtration, and DEAE-Sephadex A-50 ion exchange chromntography. The purified enzyme exhibited an optimum pH at 1.0, and an optimum temperature of 5$0^{\circ}C$. Metal ions such as Mn$^{2+}$ and $K^+$, dithiothreitol, and 2-mercaptoethanol stimulated $\beta$-galactosidase activity. Ethylenediamine tetraacetic add, 8-hydroxyquinoline, Hg$^2+$, Zn$^{2+}$, Co$^{2+}$, $Ca^{2+}$, and galactose were inhibitory. The $K_m$ and V$_{max}$ for o-nitrophenyl $\beta$-D-galactopyranoside were 1.25mM and 88.50$\mu$moles/min.mg protein, respectively. The molecular weight was estimated to be 520,000, and the amino acid composition indicated relatively high contents of glutamic acid, aspartic acid, leucine, and valine.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1581-1588
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• 2018

The growth of lactic acid bacteria (LAB) generates a high number of metabolites related to aromas and flavors in fermented dairy foods. These microbial proteases are involved in protein hydrolysis that produces necessary peptides for their growth and releases different molecules of interest, like bioactive peptides, during their activity. Each genus in particular has its own proteolytic system to hydrolyze the necessary proteins to meet its requirements. This review aims to highlight the differences between the proteolytic systems of Streptococcus thermophilus and other lactic acid bacteria (Lactococcus and Lactobacillus) since they are microorganisms that are frequently used in combination with other LAB in the elaboration of fermented dairy products. Based on genetic studies and in vitro and in vivo tests, the proteolytic system of Streptococcus thermophilus has been divided into three parts: 1) a serine proteinase linked to the cellular wall that is activated in the absence of glutamine and methionine; 2) the transport of peptides and oligopeptides, which are integrated in both the Dpp system and the Ami system, respectively; according to this, it is worth mentioning that the Ami system is able to transport peptides with up to 23 amino acids while the Opp system of Lactococcus or Lactobacillus transports chains with less than 13 amino acids; and finally, 3) peptide hydrolysis by intracellular peptidases, including a group of three exclusive of S. thermophilus capable of releasing either aromatic amino acids or peptides with aromatic amino acids.

• Food Science of Animal Resources
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• v.35 no.5
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• pp.683-691
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• 2015

The objective of this study was to identify the coccoidal bacteria present in 188 samples of fermented yaks’, mares’ and cows’ milk products collected from 12 different regions in Mongolia. Furthermore, we evaluated the fermentation properties of ten selected isolates of the predominant species, Streptococcus (S.) thermophiles, during the process of milk fermentation and subsequent storage of the resulting yoghurt at 4℃. Overall, 159 isolates were obtained from 188 samples using M17 agar. These isolates were presumed to be lactic acid bacteria based on their gram-positive and catalase-negative properties, and were identified to species level using 16S rRNA gene sequence analysis. These coccoid isolates were distributed in four genera and six species: Enterococcus (E.) durans, Enterococcus (E.) faecalis, Lactococcus (Lac.) subsp. lactis, Leuconostoc (Leuc.) lactis, Leuconostoc (Leuc.) mesenteroides. subsp. mesenteroides and S. thermophilus. Among these S. thermophilus was the most common species in most samples. From evaluation of the fermentation characteristics (viable counts, pH, titratable acidity [TA]) of ten selected S. thermophilus isolates we could identify four isolates (IMAU 20246, IMAU20764, IMAU20729 and IMAU20738) that were fast acid producers. IMAU20246 produced the highest concentrations of lactic acid and formic acid. These isolates have potential as starter cultures for yoghurt production.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.158-162
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• 2010

The gene encoding N-terminally truncated Tod polymerase ($\Delta$Tod polymerase) from Thermus thermophilus HJ6 was expressed in Escherichia coli under the control of the lambda pR and pL tandem promoters on the expression vector pJLA503. The N-terminal domain (250 amino acids) of Tod polymerase was removed without significant effect on enzyme activity and stability, while no 5'$\rightarrow$3' exonuclease activity was detected. The $\Delta$Tod polymerase was verified to possess very efficient reverse transcriptase (RT) activity in the presence of $MgCl_2$. The cDNA can also be amplified in the polymerase chain reaction (PCR) with this mutant enzyme. The $\Delta$Tod polymerase was exhibited higher activity than the Taq polymerase in a one-step RT-PCR.