• Title/Summary/Keyword: Microbial transglutaminase (MTG)

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Production of Microbial-Transglutaminase [MTG] from Streptoverticillium mobaraense

  • Wang, Hong-Wei;Kim, In-Hae;Park, Chang-Su;Lee, Jae-Hwa
    • KSBB Journal
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    • v.22 no.5
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    • pp.322-327
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    • 2007
  • Mineral salts in medium usually profoundly influence microorganism growth and protein synthesis. In order to produce microbial transglutaminase (MTG) with a high yield from Streptoverticillium mobaraense, we screened the minerals $CaCl_2,\;CoCl_2,\;FeSO_4,\;ZnSO_4,\;MnSO_4\;and\;CuSO_4$ for MTG fermentation. The results indicated that appropriate $FeSO_4$ concentrations could significantly promote cell growth and stimulate the production of MTG. With 15 mg/L of $FeSO_4$ added to medium, 58% improvements were noted in MTG productivity (2.24 U/mL). NaCl, $CaCl_2,\;and\;CoCl_2$ enhanced MTG productivity by less than 15%, and the optimal concentrations were determined as 1 g/L, 2 g/L, and 30 mg/L respectively. Furthermore, it was determined that 7.5 mg/L of $ZnSO_4$ in medium could augment MTG productivity by 20% and induce the stationary phase for MTG production to a period 24 hr earlier. This basic and novel discovery should result in the development of a good complement to the previously defined culture media for MTG fermentation.

Effect of Faba Bean Isolate and Microbial Transglutaminase on Rheological Properties of Pork Myofibrillar Protein Gel and Physicochemical and Textural Properties of Reduced-Salt, Low-Fat Pork Model Sausages

  • Geon Ho Kim;Koo Bok Chin
    • Food Science of Animal Resources
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    • v.44 no.3
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    • pp.586-606
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    • 2024
  • The study was performed to determine the effect of faba bean protein isolate (FBPI) alone or in combination with microbial transglutaminase (MTG) on the rheological properties of pork myofibrillar protein gel (MPG), and physiochemical and textural properties of reduced-salt, low-fat pork model sausages (LFMSs). The cooking yields of MPGs with MTG or FBPI alone decreased and increased, respectively. However, the combination of FBPI and MTG was similar to the control (CTL) without FBPI or MTG. Gel strength values of MPG added with both FBPI and MTG were higher than treatments with FBPI or MTG alone. The hydrophobicity values of CTL were lower than those of MPG with FBPI alone, whereas the addition of MTG decreased the hydrophobicity of MPGs. The incorporation of FBPI alone or in combination with MTG decreased sulfhydryl groups (p<0.05). Shear stress values of MPGs with MTG tended to be higher than those of non-MTG treatments at all shear rates, and the addition of FBPI into MPGs increased shear stress values. Reduced-salt (1.0%) LFMSs with FBPI alone or combined with MTG had both lower cooking loss and expressible moisture values than those of CTL and similar values to the reference sample (REF, 1.5% salt). Textural properties of reduced-salt LFMSs with FBPI or MTG were similar to those of REF. These results demonstrated that the combination of FBPI and MTG could improve the water binding capacity and textural properties of pork MPGs and LFMSs and might be suitable for application in the development of healthier meat products.

Inconsistency in the Improvements of Gel Strength in Chicken and Pork Sausages Induced by Microbial Transglutaminase

  • Kawahara, S.;Ahhmed, A.M.;Ohta, K.;Nakade, K.;Muguruma, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.8
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    • pp.1285-1291
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    • 2007
  • This research investigated variation in the improvement of the texture of chicken and pork sausages induced by microbial transglutaminase (MTG). The extractability of myofibrillar proteins from these sausages as well as the ${\varepsilon}-({\gamma}-glutamyl)$lysine (G-L) content were also investigated. MTG treatment of sausages significantly increased the breaking strength values in both meat types, especially for samples incubated at $40^{\circ}C$. However, values of the breaking strength in both meat types were increased differently. The variation in protein extractability of samples incubated at $40^{\circ}C$ for both meat types could lead to some consideration of the mechanisms and the high accessions of myosin heavy chain (MHC) to MTG. SDS-PAGE analysis showed significant changes in the density of the bands after adding MTG, especially for the pork samples in which the bands disappeared totally. The G-L content in the presence of MTG was double that in control samples of both meat types. This study suggests that the binding ability of myofibrillar proteins with MTG is strong. This leads us to suggest that MTG functions positively with different improvements in the texture of chicken and pork products that are treated mechanically, such as sausages. Variability in gel improvement level between chicken and pork sausages was observed; this resulted from the variation in meat proteins in response to MTG, as well as to the original glutamyl and lysine content.

Evaluation of Acid-treated Fish Sarcoplasmic Proteins on Physicochemical and Rheological Characteristics of Pork Myofibrillar Protein Gel Mediated by Microbial Transglutaminase

  • Hemung, Bung-Orn;Chin, Koo Bok
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.50-57
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    • 2015
  • Fish sarcoplasmic protein (SP) is currently dumped as waste from surimi industry and its recovery by practical method for being the non-meat ingredient in meat industry would be a strategy to utilize effectively the fish resource. This study was aimed to apply pH treatment for fish SP recovery and evaluated its effect on pork myofibrillar protein (MP) gel. The pH values of fish SP were changed to 3 and 12, and neutralized to pH 7 before lyophilizing the precipitated protein after centrifugation. Acid-treated fish SP (AFSP) showed about 4-fold higher recovery yield than that of alkaline-treated SP and water absorption capacity was also about 1.2-fold greater. Because of the high recovery yield and water absorption capacity, AFSP was selected to incorporate into MP with/without microbial transglutaminase (MTG). The effects of AFSP and MTG on the physicochemical and rheological characteristics of MP and MP gel were evaluated. MTG induced an increase shear stress of the MP mixture and increase the breaking force of MP gels. MP gel lightness was decreased by adding AFSP. MP gel with MTG showed higher cooking loss than that without MTG. A reduction of cooking loss was observed when the AFSP was added along with MTG, where the insoluble particles were found. Therefore, AFSP could be contributed as a water holding agent in meat protein gel.

Effects of Red Bean (Vigna angularis) Protein Isolates on Rheological Properties of Microbial Transglutaminase Mediated Pork Myofibrillar Protein Gels as Affected by Fractioning and Preheat Treatment

  • Jang, Ho Sik;Lee, Hong Chul;Chin, Koo Bok
    • Food Science of Animal Resources
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    • v.36 no.5
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    • pp.671-678
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    • 2016
  • Fractioning and/or preheating treatment on the rheological properties of myofibrillar protein (MP) gels induced by microbial transglutaminase (MTG) has been reported that they may improve the functional properties. However, the optimum condition was varied depending on the experimental factors. This study was to evaluate the effect of red bean protein isolate (RBPI) on the rheological properties of MP gels mediated by MTG as affected by modifications (fractioning: 7S-globulin of RBPI and/or preheat treatment (pre-heating; 95℃/30 min): pre-heating RBPI or pre-heating/7S-globulin). Cooking yields (CY, %) of MP gels was increased with RBPI (p<0.05), while 7S-globulin decreased the effect of RBPI (p<0.05); however, preheating treatments did not affect the CY (p>0.05). Gel strength of MP was decreased when RBPI or 7S-globulin added, while preheat treatments compensated for the negative effects of those in MP. This effect was entirely reversed by MTG treatment. Although the major band of RBPI disappeared, the preheated 7S globulin band was remained. In scanning electron microscopic (SEM) technique, the appearance of more cross-linked structures were observed when RBPI was prepared with preheating at 95℃ to improve the protein-protein interaction during gel setting of MP mixtures. Thus, the effects of RBPI and 7S-globulin as a substrate, and water and meat binder for MTG-mediated MP gels were confirmed to improve the rheological properties. However, preheat treatment of RBPI should be optimized.

Heterologous Expression of Recombinant Transglutaminase in Bacillus subtilis SCK6 with Optimized Signal Peptide and Codon, and Its Impact on Gelatin Properties

  • Wang, Shiting;Yang, Zhigang;Li, Zhenjiang;Tian, Yongqiang
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1082-1091
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    • 2020
  • Microbial transglutaminases (MTGs) are widely used in the food industry. In this study, the MTG gene of Streptomyces sp. TYQ1024 was cloned and expressed in a food-grade bacterial strain, Bacillus subtilis SCK6. Extracellular activity of the MTG after codon and signal peptide (SP Ync M) optimization was 20 times that of the pre-optimized enzyme. After purification, the molecular weight of the MTG was 38 kDa and the specific activity was 63.75 U/mg. The optimal temperature and pH for the recombinant MTG activity were 50℃ and 8.0, respectively. MTG activity increased 1.42-fold in the presence of β-ME and 1.6-fold in the presence of DTT. Moreover, 18% sodium chloride still resulted in 83% enzyme activity, which showed good salt tolerance. Cross-linking gelatin with the MTG increased the strength of gelatin 1.67 times and increased the thermal denaturation temperature from 61.8 to 75.8℃. The MTG also significantly increased the strength and thermal stability of gelatin. These characteristics demonstrated the huge commercial potential of MTG, such as for applications in salted protein foods.

Effects of pH-treated Fish Sarcoplasmic Proteins on the Functional Properties of Chicken Myofibrillar Protein Gel Mediated by Microbial Transglutaminase

  • Hemung, Bung-Orn;Chin, Koo Bok
    • Food Science of Animal Resources
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    • v.34 no.3
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    • pp.307-315
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    • 2014
  • pH adjustment would be of advantage in improving the water holding capacity of muscle proteins. The objective of this study was to evaluate the addition of fish sarcoplasmic protein (SP) solution, which was adjusted to pH 3.0 or 12.0, neutralized to pH 7.0, and lyophilized to obtain the acid- and alkaline-treated SP samples, on the functional properties of the chicken myofibrillar protein induced by microbial transglutaminase (MTG). The solubility of alkaline-treated SP was higher than that of the acid counterpart; however, those values of the two pH-treated samples were lower than that of normal SP (p<0.05). All SP solutions were mixed with myofibrillar proteins (MP) extracted from chicken breast, and incubated with MTG. The shear stresses of MP with acid- and alkaline-treated SP were higher than that of normal SP. The thermal stability of MP mixture reduced upon adding SP, regardless of the pH treatment. The breaking force of MP gels with acid-treated SP increased more than those of alkaline-treated SP, while normal SP showed the highest value. The MP gel lightness increased, but cooking loss reduced, with the addition of SP. Smooth microstructure of the gel surface was observed. These results indicated that adjusting the pH of SP improved the water holding capacity of chicken myofibrillar proteins induced by MTG.

Evaluation of Porcine Myofibrillar Protein Gel Functionality as Affected by Microbial Transglutaminase and Red Bean [Vignia angularis] Protein Isolate at Various pH Values

  • Jang, Ho Sik;Lee, Hong Chul;Chin, Koo Bok
    • Food Science of Animal Resources
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    • v.35 no.6
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    • pp.841-846
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    • 2015
  • This study was investigated to determine the effect of microbial transglutaminase (MTG) with or without red bean protein isolate (RBPI) on the porcine myofibrillar protein (MP) gel functionality at different pH values (pH 5.75-6.5). Cooking yield (CY, %), gel strength (GS, gf), differential scanning calorimetry (DSC), and scanning electron microscopy (SEM) were determined to measure gel characteristics. Since no differences were observed the interaction between 1% RBPI and pH, data were pooled. CY increased with the addition of 1% RBPI, while it was not affected by pH values. GS increased with increased pH and increased when 1% RBPI was added, regardless of pH. There were distinctive endothermic protein peaks, at 56.55 and 75.02℃ at pH 5.75, and 56.47 and 72.43℃ at pH 6.5 in DSC results, which revealed decreased temperature of the first peak with the addition of 1% RBPI and increased pH. In SEM, a more compact structure with fewer voids was shown with the addition of 1% RBPI and increased pH from 5.75 to 6.5. In addition, the three-dimensional structure was highly dense and hard at pH 6.5 when RBPI was added. These results indicated that the addition of 1% RBPI at pH 6.5 in MTG-mediated MP represent the optimum condition to attain maximum gel-formation and protein gel functionality.

Effect of Fish Sarcoplasmic Protein on Quality Attributes of No-fat Chicken Sausages Mediated by Microbial Transglutaminase

  • Hemung, Bung-Orn;Chin, Koo Bok
    • Food Science of Animal Resources
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    • v.35 no.2
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    • pp.225-231
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    • 2015
  • Fish sarcoplasmic protein (SP) obtaining from lyophilization was evaluated its effect on the qualities of the no-fat chicken sausages in the presence of microbial transglutaminase (MTG) as compared to sodium tripolyphosphate (STPP). The cooking yields of all sausage samples were not different. Expressible moisture (EM) of sausage samples was reduced by adding fish SP, while the lowest EM values were observed in sausage samples containing STPP. The pH values of sausage samples were increased with the addition of fish SP and STPP. Proximate analysis revealed that the moisture, fat, and protein contents of all samples were not different (p>0.05). Textural properties (TP), measured by texture profile analysis, showed that hardness of no-fat sausages increased upon adding fish SP. However, the highest TP values were found in sausage samples with STPP. The redness values were reduced in sausage samples with STPP, while other color values were not affected by STPP. Sensory evaluation revealed that sausages with fish SP were accepted at the higher level than that of control. However, sausage samples with STPP showed highest TP and acceptability. Thus, partial substitution of STPP by SP would be possible to reduce phosphate level in the chicken sausages.

Purification and Characterization of Transglutaminase from a Newly Isolated Streptomyces platensis YK-2 (토양 방선균 Streptomyces platensis YK-2가 생산하는 Transglutaminase의 정제 및 효소학적 특성)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.801-806
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    • 2009
  • A species producing transglutaminase (EC 2.3.2.13) was isolated from forest soil and identified as Streptomyces platensis YK-2. The transglutaminase was purified from culture broth by 50% methanol precipitation, followed by successive chromatography on DEAE-Sephadex. The yield and purification-fold was 63.4% and 2.2-fold, respectively. The purified microbial transglutaminase (MTG) migrated as a single band of approximately 45 kDa upon sodium dodecyl sulfate polyacrylamide gel eletrophoresis. The isoelectric point determined by multichambered electrofocusing was pH $6.0{\sim}7.0$. The enzyme was strongly inhibited by $Hg^{++}$, but was activated by $Cd^{++}$, $Mg^{++}$, $Mn^{++}$, $Pb^{++}$ and reducing agents such as dithiothreitol and mercaptoethanol.