• Title/Summary/Keyword: Commercial enzymes

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Roles of Carbohydrate-Binding Module (CBM) of an Endo-β-1,4-Glucanase (Cel5L) from Bacillus sp. KD1014 in Thermostability and Small-Substrate Hydrolyzing Activity

  • Lee, Jae Pil;Shin, Eun-Sun;Cho, Min Yeol;Lee, Kyung-Dong;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2036-2045
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

Effect of the Extraction Conditions on the Quality Improvement of Mussel Extracts (추출조건이 홍합 추출물의 품질에 미치는 영향)

  • An, Kyung-Hee;Kim, Jong-Gun;Ko, Soon-Nam;Kim, Woo-Jung
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.1017-1023
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    • 1999
  • Several extraction conditions of mussel were investigated for preparation of the extract as a natural shellfish seasoning. The conditions studied were extraction temperature and time, addition of sodium phosphates and citrate and hydrolysis with commercial proteolytic enzymes. The extracts were prepared by deshelling, grinding and aqueous extraction followed by centrifugation and filtration. Extraction at $90^{\circ}C$ for 40min showed the highest solids yield with less fishy and high umami taste. Among the several phosphates and citrate added, $Na_{3}PO_{4}$ and $Na_{4}P_{2}O_{7}$ at 1% level were most effective in terms of the yield and umami taste. The pH effects showed that pH 10 resulted the highest solids yield of 28% with less fishy taste. Even though the effect of enzymatic hydrolysis was not greatly different among the commercial enzymes tested, Protamex and Protease II were somewhat better than other enzymes in taste. When the mussel were extracted by the combined conditions, hydrolysis with Protamex followed by extraction at $90^{\circ}C$ for 40min with addition of $Na_{3}PO_{4}$ at pH 10, the solid yields increased up to 30% which was about 58% improvement and high intensity of umami taste and less fishy flavor.

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Effects of Microbial Inoculants on the Fermentation, Nutrient Retention, and Aerobic Stability of Barley Silage

  • Zahiroddini, H.;Baah, J.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.10
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    • pp.1429-1436
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    • 2006
  • Fermentation characteristics, nutrient retention and aerobic stability of barley silages prepared using 6 commercial inoculants were evaluated using 126 mini-silos (3-L) in a completely randomized design. Whole barley forage was chopped, wilted to 39% DM and treated with water (control, S) or one of six inoculants: A (containing Lactobacillus plantarum); B (L. plantarum and Enterococcus faecium); C (L. plantarum and Pediococcus cerevisiae); D (L. plantarum, Pediococcus pentosaceus and Propionibacterium freudenreichii, plus hydrolytic enzymes); E (Lactobacillus buchneri plus hydrolytic enzymes); F (L. buchneri and P. pentosaceus plus hydrolytic enzymes). Samples of treated forage were collected for analysis at the time of ensiling, and then 18 silos of each treatment were filled, capped and weighed. Triplicate silos were weighed and opened after 1, 3, 5, 7, 33, and 61 d. On d 61, $400{\pm}5g$ of material from each silo was placed in 1-L styrofoam containers, covered with cheesecloth and held at room temperature. Silage temperature was recorded hourly for 14 d via implanted thermocouple probes. Chemical composition of the forage at ensiling was consistent with previously reported values. At d 61, pH was lowest (p<0.01) in silage S. Ammonia-N was lower (p<0.05) in silage A than in silages S, B, E, or F. Compared to pre-ensiling values, water soluble carbohydrate concentrations were elevated in silages S, A, B, C and D, and decreased in E and F. Lactic acid concentrations were similar (p>0.10) across treatments. Acetic acid levels were highest (p<0.01) in silage E and lowest (p<0.01) in silage D. Recovery of DM was lower (p<0.01) in silage F than in silages S, A, B, C, or D. On d 61, yeasts were most numerous (p<0.01) in silage D, which was the only silage in which temperature rose more than $2^{\circ}C$ above ambient during aerobic exposure. Silage D also had the highest (p<0.01) pH and ADIN content after aerobic exposure. Lactic acid and WSC content of silage D decreased dramatically during the 14-d aerobic exposure period. Yeast counts (at d 14 of exposure) were lowest (p<0.01) in silages E and F. In general, the commercial inoculants did not appear to enhance the fermentation of barley silage to any appreciable extent in laboratory silos.

Improvement on the Quality and Functionality of Skipjack Tuna Cooking Drip Using Commercial Enzymes (효소분해에 의한 참치 자숙액의 품질 및 기능성 개선)

  • Oh, Hyeun-Seok;Kim, Jin-Soo;Kim, Hye-Suk;Jee, Seung-Joon;Lee, Jae-Hyoung;Chung, In-Kwon;Kang, Kyung-Tae;Heu, Min-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.7
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    • pp.881-888
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    • 2007
  • For the use of skipjack tuna cooking drip (STC) as a source of functional seasoning, the STC was hydrolyzed with various commercial enzymes, such as Alcalase, Flavourzyme, Neutrase and Protamex, and its hydrolysate was also investigated on the food component characteristics. The hydrolysate incubated with Alcalase for 30 min (HA30) showed 56.8% for angiotensin I converting enzyme (ACE) inhibitory activity and 1.18 for antioxidative activity, which were high or similar compared to the other enzymatic hydrolysates. There were no differences in ACE inhibitory activity and antioxidative activity among HA30, two-step enzymatic hydrolysates, and ultrafilterates (molecular weight cut off, 10 kDa). The HA30 was very stable on the digestive enzymes, such as chymotrypsin, pepsin, trypsin according to the TCA (trichloroacetic acid) soluble index. The results suggested that skipjack tuna cooking drip could be used as a source for preparing functional seasoning sauce.

Improvement on the Quality and Functionality of Red Tanner Crab Cooking Drip Using Commercial Enzymes (효소분해의 의한 붉은 대게 자숙액의 품질 및 기능성 개선)

  • Kang, Kyung-Tae;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1022-1030
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    • 2007
  • For the improvement on the quality and functionality of red tanner crab cooking drip, the preparation of hydrolysates from red crab cooking drip using commercial enzymes (Alcalase, Flavourzyme, Neutrase and Protamex) was attempted and its taste, nutritional and functional characteristics were also investigated. According to the results of heavy metal contents and proximate composition, red tanner crab cooking drip (RTCCD) could be used as a food resource. From the results of the trichloroacetic acid soluble index (TSI), angiotensin I converting enzyme (ACE) inhibiting activity and antioxidative activity, RTCCD hydrolysates incubated with Alcalase for 2 hrs was superior to the other one-step hydrolysates. There were no differences in the ACE inhibiting activity and antioxidative activity between one-step hydrolysates, which was incubated with Alcalase for 2 hrs, and two-step hydrolysates sequentially incubated with Alcalase and other enzymes. Alcalase-treated hydrolysates was similar in proximate composition and Hunter color value, while high in free amino acid content compared with crab cooking drip. Total amino acid content of Alcalase-treated hydrolysates was 11.9 g/100 mL and the major amino acids were glutamic acid (10.2%), proline (10.1%) and glycine (10.7%).

Preparation of Whelk Internal Organ Jeotgal with the Addition of Commercial Proteolytic Enzymes (상업용 단백질 가수분해 효소를 첨가한 골뱅이 내장 젓갈의 제조)

  • Oh, Jeong-Hoon;Koo, Myung-O;Lee, Kyung-Eun;Lee, Seung-Cheol
    • Korean Journal of Food Science and Technology
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    • v.34 no.4
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    • pp.570-576
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    • 2002
  • For the utilization of the by-products of whelk processing, whelk internal organ with the addition of commercial proteolytic enzymes - Flavourzyme, Neutrase, Protease NP, Prozyme - were used to make jeotgal, Korean traditional salted and fermented seafood sauce. The products were prepared at salt concentration of 25% with enzyme contents 0.05 and 0.1%. The samples were stored at $10^{\circ}C$ and the chemical properties were evaluated for 6 months. The pH in all samples were decreased from near 6.8 in the beginning stage to 6.1-6.4 in the final stage of incubation. Amino nitrogen of jeotgal increased with enzyme concentration and showed maximum value, 646 mg%, at 0.1% of Flavourzyme. Total nitrogen content was increased till four months, but rapidly decreased after that. Protein degradations of whelk internal organ during maturation of jeotgals were investigated by SDS-PAGE. The patterns of degradation were different with added enzymes.

Production of Lignocellulytic Enzymes from Spent Mushroom Compost of Pleurotus eryngii (큰느타리버섯 수확 후 배지로부터 리그닌섬유소분해효소 생산)

  • Lim, Sun-Hwa;Kim, Jong-Kun;Lee, Yun-Hae;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.3
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    • pp.152-158
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    • 2012
  • The lignocellulytic enzymes including a-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) were extracted from spent mushroom compost (SMC) of Pleurotus eryngii. Different extraction buffers and conditions were tested for optimal recovery of the enzymes. The optimum extraction was shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. ${\alpha}$-Amylase was extracted with the productivity range from 1.20 to 1.6 Unit/SMC g. Cellulase was recovered with the productivity range from 2.10 to 2.80 U/gf. ${\beta}$-glucosidase and ${\beta}$-xylosidase productivities showed lowest recovery producing 0.1 U/g and 0.02 U/g, respectively. The P. eryngii SMCs collected from three different mushroom farms showed different recovery on laccase and xylanse, cellulase. Furthermore, the water extracted SMC was compared to commercial enzymes for its industrial application in decolorization and cellulase activity.

Effects of dietary humic acid and enzymes on meat quality and fatty acid profiles of broiler chickens fed canola-based diets

  • Disetlhe, Amogelang R.P.;Marume, Upenyu;Mlambo, Victor;Hugo, Arno
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.5
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    • pp.711-720
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    • 2019
  • Objective: This study was conducted to assess the effect of potassium humate and enzymes (Xylanase+Amylase+Protease) inclusion in diets on carcass characteristics, meat quality and fatty acid profiles of broilers fed canola-based diets. Methods: Two hundred and twenty broilers randomly allotted to 5 dietary treatments: the control (commercial broiler diet); CM (17.5% canola meal inclusion); CMEnz (17.5% CM inclusion+0.3 g/kg Axtra XAP); CMPh (17.5% CM inclusion+1.5% Potassium Humate, PH); and CMEnzPh (17.5% CM inclusion+1.5% PH+0.3 g/kg Axtra XAP) were slaughtered at day 42 for assessment of carcass and meat quality parameters. Results: Diet had no effect on carcass traits apart from breast muscle weight and breast muscle index. The highest breast muscle weight was observed in broilers fed CMEnz ($487.6{\pm}17.5g$) followed by those fed the control diet ($474.37{\pm}17.5g$). Diet also had no significant dietary effect on pH, temperature, drip loss and shear force values of the breast muscle. However, diet significantly affected meat colour and water-holding capacity. Broilers in the control and CMPh groups ($52.94{\pm}0.67$ and $52.91{\pm}0.67$) had the highest (p<0.05) values for lightness ($L^*$), whilst those fed CMEnzPh had the lowest value ($47.94{\pm}0.67$). In contrast, CM group had the lowest (p<0.05) value for redness ($a^*$) with CMEnzPh group having the highest values. The proportion of polyunsaturated fatty acids (PUFAs), n-6 and n-3 fatty acids and the PUFA/saturated fatty acid ratio were increased in CM-based diets containing enzymes and humic acid. Conclusion: It can, therefore, be concluded that CM can be included in broiler diets in the presence of enzymes and humic acid with positive effects on meat quality and important fatty acids that are beneficial to the health of consumers.

Modification of the characteristics of silkworm powder by treatment with alkaline protease

  • Kim, Sung-Kuk;Jo, You-Young;Lee, Kwang-Gill;Kim, Hyun-Bok;Kim, Yong Soon;Ju, Wan-Taek;Jung, Da-Eun;Kweon, HaeYong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.1
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    • pp.30-33
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    • 2015
  • Enzymatic modification of proteins is often used to increase the biological activity of materials. Silkworm powder has been investigated as a functional food resource, but no study has been performed on its modification by commercial food enzyme. Therefore, this study aimed to determine the feasibility of such modification of silkworm powder by alkaline protease. The activity of the enzyme was confirmed using an azocasein assay. Subsequently the silkworm powder was hydrolyzed by enzymatic treatment. UV visible spectrometry showed that the supernatant of silkworm powder subjected to enzymatic treatment had a stronger absorption band than the untreated powder. SDS-PAGE electrophoresis showed that the molecular weight of silkworm powder decreased on enzymatic treatment. Thus the results indicate that commercial enzymes might be used to modify the characteristics of silkworm powder.

Optimal Extraction and Characteristics of Lignocellulytic Enzymes from Various Spent Mushroom Composts (다양한 버섯 수확 후 배지로부터 목질섬유소 분해효소의 최적 추출 및 특성)

  • Lim, Sun Hwa;Lee, Yun Hae;Kang, Hee Wan
    • The Korean Journal of Mycology
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    • v.41 no.3
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    • pp.160-166
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    • 2013
  • Recovery of ${\alpha}$-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) from spent mushroom composts (SMCs) of Pleurotus cornucopiae, Pleurotus ostreatus, Pleurotus eryngii, Hericium erinaceum, Lyophyllum ulmarium, Agrocybe cylindracea, Lentinus lepideus, and Flammulina velvtipes were investigated using different extraction buffers. The maximum recovery of the enzymes was mostly detected in SMC extracts with tap water and 0.25% Triton X-100 by shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. The xylanase (152 U/g) and laccase (8.1 U/g) activities were the highest in SMC extracts from F. velvtipes and P. eryngii. In addition, high enzymatic activities of ${\alpha}$-amylase (3.6 U/g) and cellulase (3.4 U/g) was detected in SMC extract of A. cylindracea. Futhermore, cellulase and laccase activities of SMCE from P. eryngii were compared to commercial enzymes.