• Title/Summary/Keyword: Enzymatic yield

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Enzymatic synthesis of benzyl alcohol galactoside using Escherichia coli β-galactosidase (대장균 β-galactosidase를 이용한 benzyl alcohol galactoside의 합성 연구)

  • Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.572-580
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    • 2019
  • Recently, it has been reported that benzyl alcohol (BzOH) as an additive in cosmetics, food, and medicine lead to toxicity and allergy problem. Then, to circumvent this hurdle, we carried out the synthesis of benzyl alcohol galactoside (BzO-gal). Previously, it was confirmed that BzO-gal was synthesized by transgalactosylation reaction using Escherichia coli (E. coli) ${\beta}$-galactosidase (${\beta}-gal$). Meanwhile, in this study, two peaks of BzO-gal as sodium adduct ion (m/z=293.1004) and protonated ion (m/z=271.1180) were detected in the reaction mixture by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS). In addition, the amount of ${\beta}-gal$ and BzOH concentration, temperature, pH, and lactose concentration, respectively, were optimized (${\beta}-gal$, 0.75 U/mL; BzOH, 185 mM; temperature, $40^{\circ}C$, pH, 7.5; lactose, 350 g/l). Under these optimal conditions, 185 mM BzOH was converted into about 131 mM BzO-gal, in which the conversion yield was about 72%. In the future, BzO-gal will be applicable as a substitute for BzOH as a less toxic preservative for the cosmetic, pharmaceutical, and food industries, and we are planning to investigate the characteristics of BzO-gal as a preservative.

Optimal Conditions for Phenylethanol Galactoside Synthesis using Escherichia coli β-Galactosidase (대장균 베타-갈락토시데이즈를 이용한 Phenylethanol Galactoside 합성 조건의 최적화)

  • Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.1
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    • pp.99-106
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    • 2021
  • To circumvent the skin problem from phenylethanol (PhE), we have studied on the enzymatic synthesis of phenylethanol galactoside (PhE-gal) as an alternative to PhE. Base on the previous study, we optimized the reaction conditions for PhE-gal synthesis from PhE using E. coli β-galactosidase (β-gal). The optimal amount of β-gal, PhE concentration, pH, and temperature for PhE-gal synthesis were 0.45 U/ml, 1%, 8.0, 40℃, respectively. Under these conditions, about 81.9 mM PhE was converted into about 47.4 mM PhE-gal, in which the conversion yield was about 57.9%. Meanwhile, when the reaction mixture containing PhE and PhE-gal was mixed and fractionated with water-immiscible solvent (EA or MC), it was observed that PhE-gal was distributed in water phase, and PhE was distributed in solvent phase. Additionally, PhE-gal was clearly distributed into water phase when MC was used, but PE-gal was not when EA was used. In the future, we are planning to carried out the continuing study on developing an alternative cosmetic preservative using PhE-gal.

Manufacturing Process of Glucose from Agricultural Byproducts for Feeding a Biodiesel-producing Algae (농업부산물로부터 바이오 디젤 생산용 미세조류 배양액에 첨가할 당의 생산 공정 연구)

  • Kim, Seung-Ri;Han, In-Seob
    • Journal of Appropriate Technology
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    • v.5 no.2
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    • pp.91-96
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    • 2019
  • Microalgae do not require much land and make a higher efficient oil production. However, it costs still much higher than other biodiesel resources, such as crops. Sugars charge 80% of culture media when microalgae are massively cultured in the fermenter. This study aims to develop a cost-efficient process for sugar production from Chinese cabbage byproducts. Pre-treatment with 0.25% H2SO4 was most effective when chopped cabbage was incubated 50℃/130 rpm for 24 hours. To hydrolyze cabbage cellulose, we used cellulases secreted from Trichoderma. harzianum. T. harzianum was cultured at 28℃/pH 7/130 rpm for five days. Optimal enzymatic activity of cellulase was obtained by incubating at 0.24 FPU/ml/45℃/pH 5/130 rpm for three days. In comparison to other agricultural waste, such as rice straw, green tea leaves, and palm residue, Chinese cabbage produced the highest sugar yield. We found the optimal conditions to produce sugar from Chinese cabbage byproducts as a carbon source to culture biodiesel-producing microalgae. The efficient process developed in this study helps microalgae as a sustainable alternative energy source by cost-down.

Physicochemical Characteristics of ${\beta}-Glucan$ Isolated from Barley (보리로부터 분리한 ${\beta}-Glucan$의 이화학적 특성)

  • Kim, Sung-Ran;Choi, Hee-Don;Seog, Ho-Moon;Kim, Sung-Soo;Lee, Young-Tack
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1164-1170
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    • 1999
  • The physicochemical characteristics of ${\beta}-glucan$ isolated from waxy and non-waxy barley were investigated. The hull-less waxy and non-waxy barley containing 6.5% and 5.3% of total ${\beta}-glucan$ respectively, were used as a starting material. The yield and ${\beta}-glucan$ content of crude ${\beta}-glucan$ from waxy barley was 5.54% and 62.9%, respectively, and those were higher than 3.34% and 59.2% from non-waxy barley. The crude ${\beta}-glucan$ purified with selective precipitation and enzymatic treatment to obtain the ${\beta}-glucan$ isolate of high purity (>99%). The total yield of purified ${\beta}-glucan$ from waxy and non-waxy barley was 4.46% and 2.59%, respectively. The surface appearance of the purified ${\beta}-glucan$ by scanning electron microscopy (SEM) showed randomly entangled multi-net structure of ${\beta}-glucan$ microfibrils. The melting temperature of ${\beta}-glucan$ from waxy and non-waxy barley measured by differential scanning calorimetry (DSC) was $184.6^{\circ}C$, and $180.3^{\circ}C$, respectively. DSC endotherm of ${\beta}-glucan$ solution showed 2 peaks near $68^{\circ}C$ and $84^{\circ}C$. Enthalpy of phase transition was higher in non-waxy ${\beta}-glucan$ than waxy ${\beta}-glucan$, and the intrinsic viscosity of ${\beta}-glucan$ solution from waxy barley was higher than that of non-waxy ${\beta}-glucan$. The pasting viscosity of barley starch with the purified ${\beta}-glucan$ determined by Rapid Visco-Analyzer was higher than that of barley starch without ${\beta}-glucan$, and the effect of ${\beta}-glucan$ on increasing the paste viscosity was greater in non-waxy barley starch.

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Synthesis and Functional Properties of Plastein from the Enzymatic Hydrolysates of Filefish Protein 2. General Properties and IR Spectrum of Plasteins (말쥐치육 단백질의 효소적 가수분해물을 이용한 Plastein의 합성 및 그 물성 2. Plastein의 일반적 성상과 IR Spectrum)

  • KIM Se-Kwon;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.5
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    • pp.431-440
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    • 1987
  • In order to develop a new type of food source for the effective utilization of fish protein, plastein reaction was applied to improve the functional properties of filefish protein. Plasteins were synthesized from a peptic filefish protein hydrolysate by papain, pepsin, $\alpha-chymotrypsin$ and protease(from Streptomyces griceus) under the optimum conditions of previous paper). Also, L-glutamic acid diethylester and L-leucine ethylester were incorporated into plastein during the plastein reaction by papain. And, General composition, yield, molecular weight, amino acid composition, color and IR spectrum of plasteins were measured. The protein, ash and lipid content of the plasteins were $72\~78\%,\;7.4\~11.8\%\;and\;0.3\~0.9\%$ respectively. The yield of plasteins were papain $55.0\%,\;pepsin\;47.6\%,\;\alpha-chymotrypsin\;38.3\%,\;protease\;23.6\%$, glutamic acid-incorporated plastein (Glu-Plastein) $35.0\%$, and leucine-incorporated plastein (Leu-plastein) $45.7\%$. The glutamic acid and leucine content in Glu-plastein and Leu-plastein were $38.7\%,\;41,7\%$, respectively, while the contents in the peptic filefish protein hydrolysate were $16.01\%\;and\;8.16\%$, respectively. The amino acid compositions were similar to that of the original filefish muscle protein. The major molecular weights of the peptic hydrolysate estimated by gel filteration were 2,000 and 310, and those of plasteihs were 21,000 and 4,900 for papain, 24,000 for pepsin, 18,500 for $\alpha-chymotrypsin$ 6,700 for protease, 24,000 for Glu-plastein and 17,000 for Leu-plastein. The structural changes in freeze-dried filefish meat, the FPC and hydrolysate were not observed on the IR spectrum. But plasteins showed amide I band in $1,600\~l,700cm^{-1}$ range and resulted in a strong band in $800\~850\;cm^{-1},\;700\~750\;cm^{-1}\;and\;650\~700\;cm^{-1}$. The amide I band of Glu-plastein was wider than those of other plasteins and had also a small band at $1,440\;cm^{-1}$.

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Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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Characteristics of Hot-Water Extracts from Salmon Frame as Basic Ingredients for Gomtang-like Products (열수추출 연어 Frame 엑스분의 곰탕 유사 제품 베이스로서의 특성)

  • Han, Byung-Wook;Kim, Hye-Suk;Jee, Seung-Joon;Lee, Jae-Hyoung;Kim, Hyung-Jun;Park, Shin-Ho;Ji, Seong-Gil;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1326-1333
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    • 2007
  • For preparing the basic ingredients of Gomtang-like products from the extracts of salmon frame, the extraction conditions of salmon frame were examined, and the characteristics of the extracts were compared with commercial Gomtang. Based on the crude protein, Ex-N and sensory attributes, the extractions were optimized by extracting pretreated-salmon frame in 12 times (v/w) of water for 12 hrs, before filtering with cheese cloth to yield 3 times the volume of the raw material. The concentrations of heavy metals in extracts from salmon frame were below the safety limits suggested by KFDA. The mai or amino acids were glutamic acid and aspartic acid as the free amino acids, and glycine, proline, and glutamic acid as the total amino acids. The calcium and phosphorus contents were 18.0 mg/100 mL and 33.1 mg/100 mL, respectively, and they accounted for 20% and 18% of the recommended daily allowance for mineral intake. The angiotensin I converting enzyme (ACE) inhibitory activity was improved by incubation with Flavourzyme for 4 hrs and its $IC_{50}$ was 2 mg/mL. The results above suggested that the enzymatic hydrolysates from extracts of salmon frame could be used as a basic ingredient for preparing Gomtang-like products.

Production of yuzu granules using enzyme treated yuzu pulp powder and evaluation of its physiochemical and functional characterization (유자박 식이섬유를 이용한 유자과립 제조 및 이화학적 특성조사)

  • Seong, Hyeon Jun;Lee, Bo-Bae;Kim, Duck-Hyun;Lee, Seung-Hyun;Ha, Ji-Young;Nam, Seung-Hee
    • Korean Journal of Food Science and Technology
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    • v.53 no.4
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    • pp.382-390
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    • 2021
  • In this study, solubilized yuzu pulp powder (EYP) was produced using enzyme treated yuzu pulp powder (YP) and used to manufacture yuzu granules (0-20% EYP content). The physicochemical, product stability, and functional properties of Yuzu granules were compared among five enzyme treatments. Among the five treatments, CL had the highest YP solubilization yield (48.68%). Microstructural observation of EYP using FE-SEM revealed that its surface became irregular and porous after enzymatic treatment. Compared to YP, EYP had 2 times lower insoluble dietary fibers and 3 times lower hemicellulose and cellulose content. Among the yuzu granules, IV (yuzu granules with 15% EYP) had an excellent water and oil holding capacity and flowability. IV granule had the highest narirutin and hesperidin content of 3.4 mg and 2.2 mg/g DW, respectively and the highest antioxidant (68.4%) and tyrosinase inhibitory activities (82.5%). Therefore, EYP or granule with EYP can be used as a functional component in food industry or pharmaceutical field.

Dehydration of Lactic Acid to Bio-acrylic Acid over NaY Zeolites: Effect of Calcium Promotion and KOH Treatment (NaY 제올라이트 촉매 상에서 젖산 탈수반응을 통한 바이오아크릴산 생산: Ca 함침 및 KOH 처리 영향)

  • Jichan, Kim;Sumin, Seo;Jungho, Jae
    • Clean Technology
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    • v.28 no.4
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    • pp.269-277
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    • 2022
  • With the recent development of the biological enzymatic reaction industry, lactic acid (LA) can be mass-produced from biomass sources. In particular, a catalytic process that converts LA into acrylic acid (AA) is receiving much attention because AA is used widely in the petrochemical industry as a monomer for superabsorbent polymers (SAP) and as an adhesive for displays. In the LA conversion process, NaY zeolites have been previously shown to be a high-activity catalyst, which improves AA selectivity and long-term stability. However, NaY zeolites suffer from fast deactivation due to severe coking. Therefore, the aim of this study is to modify the acid-base properties of the NaY zeolite to address this shortcoming. First, base promoters, Ca ions, were introduced to the NaY zeolites to tune their acidity and basicity via ion exchange (IE) and incipient wetness impregnation (IWI). The IWI method showed superior catalyst selectivity and stability compared to the IE method, maintaining a high AA yield of approximately 40% during the 16 h reaction. Based on the NH3- and CO2-TPD results, the calcium salts that impregnated into the NaY zeolites were proposed to exit as an oxide form mainly at the exterior surface of NaY and act as additional base sites to promote the dehydration of LA to AA. The NaY zeolites were further treated with KOH before calcium impregnation to reduce the total acidity and improve the dispersion of calcium through the mesopores formed by KOH-induced desilication. However, this KOH treatment did not lead to enhanced AA selectivity. Finally, calcium loading was increased from 1wt% to 5wt% to maximize the amount of base sites. The increased basicity improved the AA selectivity substantially to 65% at 100% conversion while maintaining high activity during a 24 h reaction. Our results suggest that controlling the basicity of the catalyst is key to obtaining high AA selectivity and high catalyst stability.

Studies on the Production of Alcohol from Woods (목재(木材)를 이용(利用)한 Alcohol 생산(生産)에 관(關)한 연구(硏究))

  • Cheong, Jin Cheol
    • Journal of Korean Society of Forest Science
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    • v.59 no.1
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    • pp.67-91
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    • 1983
  • In order to examine the alcohol production from softwoods (Pinus densiflora Sieb. et Zucc., Pinus rigida Miller, Larix leptolepis Gordon) and hardwoods (Alnus japonica Steud., Castanea crenata Sieb. et Zucc. Populus euramericana CV 214), chemical compositions were analyzed and conditions of acid hydrolysis with wood meals were established. Also strains which could remarkably decompose the cellulose were identified, and conditions of cellulase production of strains, characteristics of cellulase, and alcohol fermentation were examined. The results were summarized as follows. 1) In acid hydrolysis of wood, the high yield of reducing sugars was shown from 1.0% to 2.0% of hydrochloric acid and 2.0% of sulfuric acid. The highest yield was produced 23.4% at wood meals of Alnus japonica treated with 1.0% of hydrochloric acid. 2) The effect of raising the hydrolysis was good at $1.5kg/cm^2$, 30 times (acid/wood meal), and 45 min in treating hydrochloric acid and 30 min in treating sulfuric acid. 3) The pretreatments with concentrated sulfuric acid were more effective concentration ranged from 50% to 60% than that with hydrochloric acid and its concentration ranged from 50% to 60%. 4) The quantative analysis of sugar composition of acid hydrolysates revealed that glucose and arabinose were assayed 137.78mg and 68.24mg with Pinus densiflora, and 102.22mg and 65.89mg with Alnus janonica, respectively. Also xylose and galactose were derived. 5) The two strains of yeast which showed remarkably high alcohol productivity were Saccharomyces cerevisiae JAFM 101 and Sacch. cerevisiae var. ellipsoldeus JAFM 125. 6) The production of alcohol and the growth of yeasts were effective with the neutralization of acid hydrolysates by $CaCO_3$ and NaOH. Production of alcohol was excellent in being fermented between pH 4.5-5.5 at $30^{\circ}C$ and growth of yeasts between pH 5.0-6.0 at $24^{\circ}C$. 7) The production of alcohol was effective with the addition of 0.02% $(NH_2)_2CO$ and $(NH_4)_2SO_4$, 0.1% $KH_2PO_4$, 0.05% $MgSO_4$, 0.025% $CaCl_2$, 0.02% $MnCl_2$. Growth of yeasts was effective with 0.04-0.06% $(NH_2)_2CO$ and $(NH_4)_2SO_4$, 0.2% $K_2HPO_4$ and $K_3PO_4$, 0.05% $MgSO_4$, 0.025% $CaCl_2$, and 0.002% NaCl. 8) Among various vitamins, the production of alcohol was effective with the addition to pyridoxine and riboflavin, and the growth of yeasts with the addition to thiamin, Ca-pantothenate, and biotin. The production of aocohol was increased in 0.1% concentration of tannin and furfural, but mas decreased in above concentration. 9) In 100ml of fermented solution, alcohol and yeast were produced 2.201-2.275ml and 84-114mg for wood meals of Pinus densiflora, and 2.075-2.125ml and 104-128mg for that of Alnus japonica. Residual sugars were 0.55-0.60g and 0.60-0.65g for wood meals of Pinus densiflora and Alnus japonica, respectively, and pH varied from 3.3 to 3.6. 10) A strain of Trichoderma viride JJK. 107 was selected and identified as its having the highest activity of decomposing cellulose. 11) The highest cellulase production was good when CMCase incubated for 5 days at pH 6.0, $30^{\circ}C$ and xylanase at pH 5.0, $35^{\circ}C$. The optimum conditions of cellulase activity were proper in case of CMCase at pH 4.5, $50^{\circ}C$ and xylanase at pH 4.5, $40^{\circ}C$. 12) In fermentation with enzymatic hydrolysates, the peracetic acid treatment for delignification showed the best yields of alcohol and its ratio was effective with the addition of about 10 times. 13) The production of alcohol was excellent when wood meals and Koji of wheat bran was mixed with 10 to 8 and the 10g of wood meals of Pinus densiflora produced 2.01-2.14ml of alcohol and Alnus japonica 2.11-2.20ml.

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