• Title/Summary/Keyword: Fermentation broth

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Separation of Nattokinase from Bacillus subtilis Fermentation Broth by Expanded Bed Adsorption with Mixed-mode Adsorbent

  • Lu Miao-Hua;Lin Dong-Qiang;Wu Yuan-Chun;Yun Jun-Xian;Mei Le-He;Yao Shan-Jing
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.128-135
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    • 2005
  • Mixed-mode hydrophobic/ionic matrices exhibit a salt-tolerant property for adsorbing target protein from high-ionic strength feedstock, which allows the application of undiluted feedstock via an expanded bed process. In the present work, a new type of mixed-mode adsorbent designed for expanded bed adsorption, Fastline $PRO^{\circledR}$, was challenged for the capture of nattokinase from the high ionic fermentation broth of Bacillus subtilis. Two important factors, pH and ion concentration, were investigated with regard to the performance of nattokinase ad-sorption. Under initial fermentation broth conditions (pH 6.6 and conductivity of 10 mS/cm) the adsorption capacity of nattokinase with Fastline PRO was high, with a maximum capacity of 5,350 U/mL adsorbent. The elution behaviors were investigated using packed bed adsorption experiments, which demonstrated that the effective desorption of nattokinase could be achieved by effecting a pH of 9.5. The biomass pulse response experiments were carried out in order to evaluate the biomass/adsorbent interactions between Bacillus subtilis cells and Fastline PRO, and to demonstrate a stable expanded bed in the feedstock containing Bacillus subtilis cells. Finally, an EBA process, utilizing mixed-mode Fastline PRO adsorbent, was optimized to capture nattokinase directly from the fermentation broth. The purification factor reached 12.3, thereby demonstrating the advantages of the mixed-mode EBA in enzyme separation.

Recovery of Lactic Acid from Fermentation Broth Using Precipitation and Reactive Distillation (발효액으로부터 침전과 반응증류를 이용한 젖산의 회수)

  • Park, Suk-Chan;Lee, Sang-Mok;Kim, Young-Jun;Kim, Woo-Sik;Koo, Yoon-Mo
    • KSBB Journal
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    • v.21 no.3
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    • pp.199-203
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    • 2006
  • Precipitation and reactive distillation were employed to recover lactic acid from fermentation broth. Lime was initially added to fermentation broth in order to convert soluble lactic acid to an insoluble calcium lactate form. Drowning-out crystallization was used to decrease the solubility of calcium lactate by adding ethanol as a co-precipitant. In the ideal solution of organic acids as well as fermentation broth, precipitation experiments were performed with varying amounts of ethanol. Precipitation process was followed by reactive distillation. Carboxylate salts formed in the previous precipitation process were mixed with carbon dioxide and triethylamine to precipitate as calcium carbonate. The remaining liquid was distilled for 1 hr at different temperatures. Triethylamine and water were recovered from the top of the distiller, while organic acids, inducing lactic acid as a main component remained in feeding bottle. The yield of recovered lactic acid was 67.5% with the purity of 99.7%.

Changes in Enzymatic Activities during Eoyukjang Fermentation (어육장 발효 시 생성되는 효소의 활성 변화)

  • Ham, Soo-Nam;Kim, Sang-Woo;Lee, Jae Hwan;Chang, Pahn-Shick
    • Korean Journal of Food Science and Technology
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    • v.40 no.3
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    • pp.251-256
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    • 2008
  • Eoyukjang is a traditional sauce-type of Korean food that is similar to a soybean sauce made from fermented soybeans, and it is produced from a fermented mixture of sea foods, meats, and meju (soybean paste). This study examined periodical changes in the enzymatic activities of ${\alpha}$-amylase, esterase, ${\beta}$-glucosidase, protease, lipase, and lipoxygenase within the culture broth and solids of eoyukjang during 1 year of fermentation. The eoyukjang solids had 234-532% higher protein content than the culture broth. The specific activities of ${\alpha}$-amylase, esterase, ${\beta}$-glucosidase, and protease increased in both the culture broth and solids. Particularly, in the culture broth, ${\alpha}$-amylase, esterase, ${\beta}$-glucosidase, and protease activities rapidly increased (3- to 8-fold) until 10 months of fermentation, and then drastically decreased. However, the activities of lipase and lipoxygenase in both the culture broth and solids were less than 0.05 unit/mg of protein, respectively, throughout fermentation; thus, their activity levels were low and changed little over the 12 months. Overall, while the solids had higher protein content than the culture broth, the broth had greater enzyme activity levels during eoyukjang preparation.

Characteristics of Fermented Fruit and Vegetable Mixed Broth Using by Bacteriocin-producing Lactic Acid Bacteria and Yeast (박테리오신 생성 젖산균과 효모를 이용한 과채발효액의 특성)

  • Jung, Dong-Sun;Lee, Young-Kyung;Lim, Kyung-Wha
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1358-1364
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    • 2000
  • A mixed culture of bacteriocin-producing lactic acid bacteria and yeast was used to prepare unique fermented fruit and vegetable beverage which contains bacteriocin. Fruit and vegetable broth fermented by Lactococcus lactis 11454 showed inhibitory activity against foodborne pathogens such as Listeria monocytogens and Staphylococcus aureus, and Streptococcus mutans which is a major causative agent for dental caries. Bacteriocin was detected at the end of fermentation of fruit and vegetables and its antimicrobial activity was stable for 5 weeks during storage at $4^{\circ}C$. These results suggest that the presence of bacteriocin in beverages during fermentation and storage will provide a safeguard against foodborne pathogens and spoilage bacteria. To improve flavor of the fermented broth, post-fermentation of the fermented broth was carried out at $4^{\circ}C$ by using yeast isolated from raw material mixture. Total acidity of the post-fermented broth was slightly decreased, but significant increase in the concentration of succinic acid was observed in the post-fermented broth. It was also observed that bitter and disagreeable taste compounds such as phthalates were decreased, and mild acidic and fruity flavor esters and alcohols were increased by the post-fermentation of the fermented broth with yeast.

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Making of Dongchimi Naengmyeun Broth Which Has Enhanced Antioxidant Activity Using Purple Sweet Potato (자색고구마를 사용한 항산화 활성이 강화된 동치미 냉면육수 제조)

  • Seo, Weon-Taek;Kim, Han-Gon;Lee, Jin-Sung;Cho, Kye-Man
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.143-150
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    • 2011
  • Lactic fermented dongchimi naengmyeun broth which has enhanced antioxidative activity was developed. During lactic fermentation of dongchimi naengmyeun broth at $25^{\circ}C$, changes of total lactic acid bacteria, pH, acidity, soluble phenolics, and anthocyanins were investigated. After 72 h of fermentation, the stronger antioxidant activities were observed in dongchimi naengmyeun broth supplemented with purple sweet potato than those of control dongchimi naengmyeun broth which showing 96.80% in DPPH radical scavenging activity, 100.82% in $ABTs^{+{\cdot}}$ scavenging activity, 7.77 in reducing power, and 6.89 in ferric reducing/antioxidant power, respectively. These high antioxidant activities related with higher contents of soluble phenolics and anthocyanins in dongchimi naengmyeun broth supplemented with purple sweet potato. The results suggest that the making of functional dongchimi naengmyeun broth by using high soluble phenolics and anthocyanins supplements such as purple sweet potato powder was possible.

Isolation and Properties of Amino Acid Antimetabolite from Streptomyces sp. YS-943 (Streptomyces sp. YS-943균주가 생산하는 아미노산 대사길항물질의 정제와 성상)

  • 유성재;박부길
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.81-86
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    • 1995
  • A Streptomyces strain YS-943, which produced amino acid antimetabolite, was isolated from soil. During the course of screening for new amino acid antimetabolites from the culture broth of Actinomycetes, we found that the strain produced a substance active against Gram-positive bacteria and its activity was reversed by L-methionine and L-histidine on the synthetic minimal agar medium in the culture broth.The morphological and cultural characteristics serve to identify the producing organism strain YS-943 as the genus Streptomyces. Fermentation was carried out in the synthetic medium at 28$\CIRC$C for 48 hours. The fermentation yield reached about 12 mg per liter of the broth. The YS-943 substance was obtained as white powder, mp 194$\CIRC$C and has the molecular formular of C$_{4}$H$_{8}$N$_{2}$O$_{4}$. Its structure was determined to be o-carbamyl-D-serine by spectroscopic data. It is active against some Gram-positive bacteria and reversed by L-methionine and L-histidine.

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나노여과를 이용한 발효유산 정제에서의 hardness $(Mg^{2+},\;Ca^{2+})$ 제거

  • Gang, Sang-Hyeon;Lee, Eun-Gyo;Jeong, Yong-Ho;Jang, Yong-Geun
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.489-492
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    • 2000
  • The rejection of hardness in a lactate fermentation broth was investigated by NTR-729 nanofiltration membrane. Firstly, a method for determining membrane surface charge was proposed on the basis of the rejection data for NaCl, $Na_2SO_4$, $MgCl_2$, $MgSO_4$ solutions. It was found that the NTR-729 membrane was negatively charged. The rejection of $Mg^{2+}$ ions in model solution of lactate was over 80% with lactate concentrations. But the rejection of $Mg^{2+}$ ions in fermentation broth of lactate was about 40 %. This results was caused by anions paired with $Mg^{2+}$ ions. It was found that the repulsive force of monovalent anions with membrane was almost few through an experiment of a mixed solution of NaCl/ $Na_2SO_4$. So the rejection of $Mg^{2+}$ ion in a fermentation broth was lower than that in a model solution because $Mg^{2+}$ ions permeated to the membrane with various monovalent anions.

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Recovery of Lactic Acid from Fermentation Broth by the Two-Stage Process of Nanofiltration and Water-Splitting Electrodialysis

  • Lee, Eun-Gyo;Kang, Sang-Hyeon;Kim, Hyun-Han;Chang, Yong-Keun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.313-318
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    • 2006
  • A two-stage process of nanofiltration and water-splitting electrodialysis was investigated for lactic acid recovery from fermentation broth. In this process, sodium lactate is isolated from fermentation broth in the first stage of nanofiltration by using an NTR-729HF membrane, and then is converted to lactic acid in the second stage by water-splitting electrodialysis. To determine the optimal operating conditions for nanofiltration, the effects of pressure, lactate concentration, pH, and known added impurities were studied. Lactate rejection was less than 5%, magnesium rejection approximated 45%, and calcium rejection was at 40%. In subsequent water-splitting electrodialysis, both the sodium lactate conversion to lactic acid and sodium hydroxide recovery, were about 95%, with a power requirement of $0.9{\sim}1.0\; kWh$ per kg of lactate.

Determination of Nebramycin Factor 2,4,5,5',6 and Kanamycin A in Fermentation Broth of Streptoalloteichus hindustanus ATCC 31218 Mutant Using 2,4-Dinitrofluorobenzene(DNFB) as a Derivatizing Agent by High Performance Liquid Chromatography (HPLC법에 의한 2,4-디니트로플루오로벤젠을 유도체화제로 한 Streptoalloteichus hindustanus ATCC 31218 변이균의 배양액 중 네브라마이신 펙터 2,4,5,5',6, 가나마이신 A 분석)

  • 박영근;박명용;김승철;양호길
    • YAKHAK HOEJI
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    • v.37 no.1
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    • pp.1-8
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    • 1993
  • A procedure for the high-performance liquid chromatographic determination of Nebramycin factors in fermentation broth of Streptoalloteichus hindustanus ATCC 31218 mutant was investigated using pre-column derivatization and LTV detection. The method is based on pre-column derivatization of Nebramycin factors with 2,4-dinitrofluorobenzene(DNFB) in the presence of Tris (hydroxymethyl)aminoethane. The chromatographic separation of derivatives of Nebramycin factors and unknown impurities is achieved using reversed-phase column (NOVA-PAK $C_{18}$, Waters Co.) and AcCN : H$_{2}$O : AcOH (53.0:46.5:0.5) as a mobile phase. The mixture of these derivatives were separated within 35 minutes and the optimum wavelength($\lambda_{max}$ ) of the UV detector was 353 nm. The linearity of response for derivatives of Nebramycin factors is demonstrated for concentrations up to 500 $\mu\textrm{g}$/ml and the relative standard deviation is less than 0.79%. Detection limit was 1.67 ng for the 10 $\mu\textrm{l}$ sample volume employed.

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Construction of Amylolytic Industrial Brewing Yeast Strain with High Glutathione Content for Manufacturing Beer with Improved Anti-Staling Capability and Flavor

  • Wang, Jin-Jing;Wang, Zhao-Yue;He, Xiu-Ping;Zhang, Bo-Run
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1539-1545
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    • 2010
  • In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.