• Title/Summary/Keyword: Two-phase fermentation

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Cell Fusion Between Xylose Fermenting Yeast and Xylanase Secreting Yeast (Xylanase 분비효모와 Xylose 발효효모의 Protoplast Fusion)

  • 김남순;배명애;서정훈
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.88-93
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    • 1989
  • To improve a new yeast strain capable of converting xylan to ethanol directly, we tried protoplast fusion between xylose fermenting yeast (Candida sp. X-6-41) and xylan assimilating yeast (Crypto-coccus sp. XB-33), finally selected the most promising two fusants (XFU-1 and XFU-2). As the optimum conditions for protoplast formation, the yeast cells were cultured to exponential phase in YPD and YPX containing 0.6M KCI, respectively, and then treated with zymolyase (0.25mg/$m\ell$), cellulase(4mg/$m\ell$) and 100mM 2-mercaptoethanol at pH 8 and 3$0^{\circ}C$. The protoplasts of parental auxotrophs were fused in the presence of 20mM CaCl$_2$and 40% polyethylene glycol(M.W.4000). The physiological and morphological characteristics of the fusants, such as assimilation of carbon sources, cell size, growth rate, xylanase activity and xylan fermentation ability were investigated. Xylanase activity of fusants that cultured in chemically minimal medium was higher than that of fusants that cultured in completed medium, because xylanase producing activity of xylose fermenting yeast(X-6-41) was inhibited by isoleucine.

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Efficient and Cost-Reduced Glucoamylase Fed-Batch Production with Alternative Carbon Sources

  • Luo, Hongzhen;Liu, Han;He, Zhenni;Zhou, Cong;Shi, Zhongping
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.185-195
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    • 2015
  • Glucoamylase is an important industrial enzyme. Glucoamylase production by industrial Aspergillus niger strain featured with two major problems: (i) empirical substrate feeding methods deteriorating the fermentation performance; and (ii) the high raw materials cost limiting the economics of the glucoamylase product with delegated specification. In this study, we first proposed a novel three-stage varied-rate substrate feeding strategy for efficient glucoamylase production in a 5 L bioreactor using the standard feeding medium, by comparing the changing patterns of the important physiological parameters such as DO, OUR, RQ, etc., when using different substrate feeding strategies. With this strategy, the glucoamylase activity and productivity reached higher levels of 11,000 U/ml and 84.6 U/ml/h, respectively. The performance enhancement in this case was beneficial from the following results: DO and OUR could be controlled at the higher levels (30%, 43.83 mmol/l/h), while RQ was maintained at a stable/lower level of 0.60 simultaneously throughout the fed-batch phase. Based on this three-stage varied-rate substrate feeding strategy, we further evaluated the economics of using alternative carbon sources, attempting to reduce the raw materials cost. The results revealed that cornstarch hydrolysate could be considered as the best carbon source to replace the standard and expensive feeding medium. In this case, the production cost of the glucoamylase with delegated specification (5,000 U/ml) could be saved by more than 61% while the product quality be ensured simultaneously. The proposed strategy showed application potential in improving the economics of industrial glucoamylase production.

Potential Antioxidant Peptides in Rice Wine

  • Rhee, Sook-Jong;Lee, Chung-Yung J.;Kim, Mi-Ryung;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.715-721
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    • 2004
  • Many food protein hydrolysates have been shown to have antioxidant activities, and recent research focuses on low molecular peptides produced during hydrolysis of food protein. Korean rice wine contains about 60-70% of protein at dry base and originates from raw materials. It has been suggested that the protein is transformed into low molecular weight peptides, and have antioxidant activity during fermentation. The objectives of this study were to evaluate the antioxidant activity of the pre-purified and purified peptides found in Korean rice wine and to identify the responsible peptides. The wine extract of Samhaeju, a traditional Korean rice wine made by low temperature fermentation, was evaporated at $35^{\circ}C$. The two methods employed in the evaluation of antioxidant activity were the DPPH radical scavenging method and the beta-carotene bleaching test. The pre-purified samples showed 808 AAC (Antioxidant Activity Coefficient) and 56.5% AOA (Antioxidant Activity), which were higher than $\alpha$-tocopherol (572 AAC and 78% AOA). The rice wine extract was separated by reversed-phase HPLC. The protective effect of the four most antioxidant active fractions were tested for t-butyl hydroperoxide induced oxidation of healthy human erythrocytes and the byproduct was determined by malondialdehyde formation. Fraction No.5 showed 35% lower MDA concentration as compared to the control. The peptides were further purified using consecutive chromatographic methods and 4 antioxidant peptides were isolated. The amino acid sequences of the peptides were identified as Ile-His-His, Val- Val-His(Asn), Leu-Val-Pro, and Leu(Val)-Lys-Arg-Pro. The AAC value of the synthetic form of the identified peptides was the highest for Ile-His-His.

Investigation on the Microbiological and Biochemical Properties of Kimchi in the Solid-state Model System Designed for Fermented Sausages

  • Lee, Joo-Yeon
    • Food Science of Animal Resources
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    • v.30 no.2
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    • pp.236-242
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    • 2010
  • The objective of this study was to investigate the potential of the application of kimchi LAB as starter culture in the production of fermented sausages. For this, the solid-state model media composed to simulate the substantial conditions of meat mixtures were fermented for 120 h after the treatment with different concentrations of kimchi (0.5, 1.0, 1.5, 3.0, and 5.0%) and lyophilized kimchi-powder (0.2 % and 0.5%). During the fermentation period, the growth of total viable cells and LAB, and the changes of pH and titratable acidity were investigated. The initial LAB counts ranged from 7.18 to 8.34 Log CFU/ mL for kimchi media and from 6.93 to 6.94 Log CFU/mL for kimchi-powder media depending on the added concentrations. The kimchi LAB in this study were not influenced by the immobilized condition for their adaptation and growth by showing no lag phase and thus acted similar as in the submerged medium. The initially increased counts reached around 9 Log CFU/ mL in 12 h independent of the concentrations of a ded kimchi. However, the growth and metabolic activity of kimchi-powder LAB were influenced by the immobilized condition. Supposedly, as the nutrient supply in solid-state depended solely on diffusion, these differences in the souring properties were caused by the LAB topography in the medium matrix. Nevertheless, the differences in the numbers of LAB between two media were less than 0.5 Log units and the pH drop in the solidstate batches was quite rapid and reached low values. Therefore, it can be assumed that kimchi and kimchi-powder LAB showed the utility as the substitute of commercial starter culture even without a rehydrating pretreatment.

Mcl-PHAs Produced by Pseudomonas sp. Gl01 Using Fed-Batch Cultivation with Waste Rapeseed Oil as Carbon Source

  • Mozejko, Justyna;Wilke, Andreas;Przybylek, Grzegorz;Ciesielski, Slawomir
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.371-377
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    • 2012
  • The present study describes medium-chain-length polyhydroxyalkanoates (mcl-PHAs) production by the Pseudomonas Gl01 strain isolated from mixed microbial communities utilized for PHAs synthesis. A two-step fed-batch fermentation was conducted with glucose and waste rapeseed oil as the main carbon source for obtaining cell growth and mcl-PHAs accumulation, respectively. The results show that the Pseudomonas Gl01 strain is capable of growing and accumulating mcl-PHAs using a waste oily carbon source. The biomass value reached 3.0 g/l of CDW with 20% of PHAs content within 48 h of cultivation. The polymer was purified from lyophilized cells and analyzed by gas chromatography (GC). The results revealed that the monomeric composition of the obtained polyesters depended on the available substrate. When glucose was used in the growth phase, 3-hydroxyundecanoate and 3-hydroxydodecanoate were found in the polymer composition, whereas in the PHAs-accumulating stage, the Pseudomonas Gl01 strain synthesized mcl-PHAs consisting mainly of 3-hydroxyoctanoate and 3-hydroxydecanoate. The transcriptional analysis using reverse-transcription real-time PCR reaction revealed that the phaC1 gene could be transcribed simultaneously to the phaZ gene.

Protoplast fusion between saccharomyces cerevisiae and candida cariosilignicola (Saccharomyces cerevisiae와 candida cariosilignicola사이의 세포융합에 관한 연구)

  • 이재동;임하선
    • Korean Journal of Microbiology
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    • v.26 no.1
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    • pp.37-43
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    • 1988
  • This research was focused on investigation of the condition for protoplast formation and regeneration of protoplast fusion between Saccharomyces cerevisiae which has fermentation ability and Candida cariosilignicola which can grow at high temperature and utilize methanol. The results obtained were as follows; The highest production was collected in exponential growth phase. Ninety-nine% protoplast formation of C. cariosilignicola was obtained in glycin-NaOH buffer (pH10.0) containing Zymolyase 0.5mg/ml at $35^{\circ}C$ for 1hr incubation. The highest regeneration was produced when protoplast wuwpension containing 0.5% soft agar in buffered 50mM $CaCl_{2}$ was poured as a soft overlay onto 2% agar plates. Equal amuont of protoplast suspension of two strains was mixed and centrifuged. The subsequent pellet was added to 2ml of 35% polyethylene glycol (MW 4,000) containing 50mM $CaCl_{2}$, and incubated at $30^{\circ}C$ for 10min. Then 0.1ml of the suspension of aggregated protoplast was immediately covered with minimal medium and incubated at $40^{\circ}C$ for 5-7 days. As results, $SC_{1}$, $SC_{2}$, and $SC_{3}$ fusants were obtained. The physiological characteristics of fusants produced by protoplast fusion were; $SC_{1}$, and $SC_{2}$ utilized maltose, galactose, methanol, potassium nitrate. $SC_{3}$ utilized all the above materials except galactose.

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Evaluation of liquid and powdered forms of polyclonal antibody preparation against Streptococcus bovis and Fusobacterium necrophorum in cattle adapted or not adapted to highly fermentable carbohydrate diets

  • Cassiano, Eduardo Cuellar Orlandi;Perna, Flavio Junior;Barros, Tarley Araujo;Marino, Carolina Tobias;Pacheco, Rodrigo Dias Lauritano;Ferreira, Fernanda Altieri;Millen, Danilo Domingues;Martins, Mauricio Furlan;Pugine, Silvana Marina Piccoli;de Melo, Mariza Pires;Beauchemin, Karen Ann;Meyer, Paula Marques;Arrigoni, Mario de Beni;Rodrigues, Paulo Henrique Mazza
    • Animal Bioscience
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    • v.34 no.1
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    • pp.74-84
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    • 2021
  • Objective: Feed additives that modify rumen fermentation can be used to prevent metabolic disturbances such as acidosis and optimize beef cattle production. The study evaluated the effects of liquid and powdered forms of polyclonal antibody preparation (PAP) against Streptococcus bovis and Fusobacterium necrophorum on rumen fermentation parameters in ruminally cannulated non-lactating dairy cows that were adapted or unadapted to a high concentrate diet. Methods: A double 3×3 Latin square design was used with three PAP treatments (control, powdered, and liquid PAP) and two adaptation protocols (adapted, unadapted; applied to the square). Adapted animals were transitioned for 2 weeks from an all-forage to an 80% concentrate diet, while unadapted animals were switched abruptly. Results: Interactions between sampling time and adaptation were observed; 12 h after feeding, the adapted group had lower ruminal pH and greater total short chain fatty acid concentrations than the unadapted group, while the opposite was observed after 24 h. Acetate:propionate ratio, molar proportion of butyrate and ammonia nitrogen concentration were generally greater in adapted than unadapted cattle up to 36 h after feeding. Adaptation promoted 3.5 times the number of Entodinium protozoa but copy numbers of Streptococcus bovis and Fibrobacter succinogens genes in rumen fluid were not affected. However, neither liquid nor powdered forms of PAP altered rumen acidosis variables in adapted or unadapted animals. Conclusion: Adaptation of cattle to highly fermentable carbohydrate diets promoted a more stable ruminal environment, but PAP was not effective in this study in which no animal experienced acute or sub-acute rumen acidosis.

Strain Selection and Optimization of Mixed Culture Conditions for Lactobacillus pentosus K1-23 with Antibacterial Activity and Aureobasidium pullulans NRRL 58012 Producing Immune-Enhancing β-Glucan

  • Sekar, Ashokkumar;Kim, Myoungjin;Jeong, Hyeong Chul;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.697-706
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    • 2018
  • Lactobacillus pentosus K1-23 was selected from among 25 lactic acid bacterial strains owing to its high inhibitory activity against several pathogenic bacteria, including Escherichia coli, Salmonella typhimurium, S. gallinarum, Staphylococcus aureus, Pseudomonas aeruginosa, Clostridium perfringens, and Listeria monocytogenes. Additionally, among 13 strains of Aureobasidium spp., A. pullulans NRRL 58012 was shown to produce the highest amount of ${\beta}$-glucan ($15.45{\pm}0.07%$) and was selected. Next, the optimal conditions for a solid-phase mixed culture with these two different microorganisms (one bacterium and one yeast) were determined. The optimal inoculum sizes for L. pentosus and A. pullulans were 1% and 5%, respectively. The appropriate inoculation time for L. pentosus K1-23 was 3 days after the inoculation of A. pullulans to initiate fermentation. The addition of 0.5% corn steep powder and 0.1% $FeSO_4$ to the basal medium resulted in the increased production of lactic acid bacterial cells and ${\beta}$-glucan. The following optimal conditions for solid-phase mixed culture were also statistically determined by using the response surface method: $37.84^{\circ}C$, pH 5.25, moisture content of 60.82%, and culture time of 6.08 days for L. pentosus; and $24.11^{\circ}C$, pH 5.65, moisture content of 60.08%, and culture time of 5.71 days for A. pullulans. Using the predicted optimal conditions, the experimental production values of L. pentosus cells and ${\beta}$-glucan were $3.15{\pm}0.10{\times}10^8CFU/g$ and $13.41{\pm}0.04%$, respectively. This mixed culture may function as a highly efficient antibiotic substitute based on the combined action of its anti-pathogenic bacterial and immune-enhancing activities.

Screening of Lactic Acid Bacteria as Starter Culture for Making Fermented Sausage (발효 소시지 제조를 위한 기능성 유산균의 선발)

  • Han, Soo-Min;Kim, Young-Joo;Lee, Hong-Chul;Chin, Koo-Bok;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.26 no.4
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    • pp.511-516
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    • 2006
  • The objectives of this study was to compare the probiotic characteristics of lactic acid bacteria (LAB) for their ability to assimilate cholesterol, production of bacteriocin, inhibition of angiotensin I-converting enzyme (ACE), and viability under artificial gastrointestinal fluids. Among tested lactic acid bacteria, L167 strain exhibited the highest ACE inhibitory activity (58.75%). The production of ACE inhibitory peptide derived from fermented milk by L167 strain started at the beginning of stationary phase with maximum activity occurring late of the stationary phase. The highest ACE inhibitory activity was observed at 20 h in 10% skim milk medium. L155 strain exhibited cholesterol assimilation activity compared with probiotic strains such as Lactobacillus acidophilus ATCC 43121. With addition of bacteriocin culture, viable cells of Staphylococcus aureus in fermented sausage were slightly decreased during storage. Among selected strains of LAB, 3 strains weve identified as L. plantarum (L155, L165, L167), and two strains were identified as Pediococcus damnosus (L12) and L. paracasei ssp. paracasei (P113) by use of API carbohydrate fermentation pattern and physiological tests.

Beet pulp as soluble fiber source and dietary energy levels for growing pigs under heat stress

  • Yo Han Choi;Ye Jin Min;Da Yeon Jeon;Hyun Ju Jin;Yong Dae Jeong;Hyun Ju Park;Abdolreza Hosseindoust;Sang Hun Ha;Jun Young Mun;Jin Soo Kim;Jo Eun Kim
    • Journal of Animal Science and Technology
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    • v.65 no.5
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    • pp.989-1001
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    • 2023
  • The study evaluated the effects of dietary fiber and energy levels administered during two growing periods (d 0-28 and d 29-56) for pigs exposed to a high temperature. A total of 96 growing pigs were used in six treatments as: Two treatments in thermoneutral temperature (21℃-24℃) with dietary energy of 3,300 and the inclusion of high or low fiber, two treatments in heat stress (30℃-34℃) with dietary energy of 3,300 and the inclusion of high or low fiber, and two treatments in heat stress with dietary energy of 3,450 and the inclusion of high or low fiber. Among standard energy level treatments, heat-stressed pigs showed lower average daily gain (ADG), feed intake, digestibility of dry matter, gross energy, crude protein, and crude fiber in phases 1 and 2. Moreover, higher concentrations of acetate, propionate, butyrate, and total short-chain fatty acid (SCFA) in feces were shown in pigs fed high fiber diets. There was a negative interaction between dietary fiber and energy for the fecal concentration of isobutyrate in phase 1 and valerate in phase 2. Pigs in heat stress treatments showed a higher rectal temperature, respiratory rate, hair cortisol, plasma zonulin, and fecal lipocalin-2. Among heat stress treatments, the overall ADG was increased in pigs fed high fiber. Pigs fed high dietary fiber showed a greater concentration of acetate, propionate, butyrate, and total SCFA. High fiber treatments decreased plasma zonulin. In conclusion, the inclusion of beet pulp, soluble fiber, at the level of 4% looks necessary in pigs diet during heat stress.