• Title/Summary/Keyword: alkaline fermentation

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Optimization of Bio-based Succinic Acid Production from Hardwood Using the Two Stage pretreatments

  • Jung, Ji Young;Jo, Jong Soo;Kim, Young Wun;Yoon, Byeng Tae;Kim, Choon Gil;Yang, Jae Kyung
    • Journal of the Korean Wood Science and Technology
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    • v.41 no.2
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    • pp.111-122
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    • 2013
  • The steam explosion-chemical pretreatment is a more effective wood pretreatment technique than the conventional physical pretreatment by accelerating reactions during the pretreatment process. In this paper, two-stage pretreatment processes of hardwood were investigated for its enzymatic hydrolysis and the succinic acid yield from the pretreated solid. The first stage pretreatment was performed under conditions of low severity to optimize the amount of solid recovery. In the second stage pretreatment washed solid material from the first stage pretreatment step was impregnated again with chemical (alkaline or chlorine-based chemicals) to remove a portion of the lignin, and to make the cellulose more accessible to enzymatic attack. The effects of pretreatment were assessed by enzymatic hydrolysis and fermentation, after the two stage pretreatments. Maximum succinic acid yield (16.1 g $L^{-1}$ and 77.5%) was obtained when the two stage pretreatments were performed at steam explosion -3% KOH.

A High Viscosity of Curdlan at Alkaline pH Increases Segregational Resistance of Concrete (염기성 pH에서의 고점도 커들란에 의한 콘크리트의 재료분리 억제 효과 증진)

  • 이인영;김선원;이중헌;김미경;조인성;박영훈
    • KSBB Journal
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    • v.14 no.1
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    • pp.114-118
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    • 1999
  • In order to use a polysaccharide, curdlan, as a concrete admixture, we first developed a pilot-scale fermentation process for the mass production of curdlan. We also examined the rheological properties of curdlan, and tested how well the curdlan obtained in this work increased the segregational resistance of the cement slurry. Fermentation was performed in a 300-liter fermenter equipped with 3 disk-turbine impellers. Since curdlan production is stimulated under nitrogen-limiting conditions, the culture pH was shifted from the optimal pH for cell growth (pH 7.0) to the optimal pH for curdlan production (pH 5.5) at the onset of ammonium exhaustion. We obtained a curdlan production of 65 g/L in 120 hr batch cultivation of Agrobacterium species. The insoluble curdlan at the final stage of fermentation was readily harvested by centrifugation together with the cells. The freeze-dried sample contained 78% (w/w) of curdlan. The solubility and viscosity of the curdlan increased with the increase of the solution pH, which enhances the viscosity of concrete since the pH of concrete is extremely high (pH 13.0). Test results of the curdlan as a concrete admixture with cement slurry demonstrated that it prohibits the leakage of water. In conclusion, this work certifies and enlarges curdlan's industrial potential as a concrete admixture.

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Preparation of Accelerated Salt-fermented Anchovy Sauce Added with Shrimp Byproducts (새우가공부산물을 이용한 속성 멸치액젓의 제조)

  • Kim, Hye-Suk;Yang, Soo-Kyeong;Park, Chan-Ho;Han, Byung-Wook;Kang, Kyung-Tae;Ji, Seung-Gil;Sye, Youn-Eon;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1265-1273
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    • 2005
  • The purpose of this study was to prepare accelerated salt-fermented anchovy sauce using a shrimp processing byproducts (head, shell and tail) as a fermenting accelerator, and to investigate its physicochemical and enzymatic properties. Four types of sauces were prepared with 0, 10, 20, and 30$\%$ (w/w) addition of shrimp byproduct and fermented at 24$\pm$2$^{\circ}C$ for 360 days. During fermentation, all four type sauces decreased moisture content (67.5$\%$68.0$\%$ to 64.0$\∼$64.8$\%$) and pH (5.52$\∼$7.10 to 5.03$\∼$6.58), but showed increase in their crude protein (7.0$\∼$8.2 to 10.8$\%$) and volatile basic nitrogen contents (40$\∼$75 to 180$\∼$200 mg/100 g of sauce). The ratio of amino nitrogen to total nitrogen contents of control (0$\%$) and sauce with 10$\%$ shrimp byproducts (10$\%$ sauce) were maximized at 270 days, whereas 20$ \% $ and 30$\%$ added sauces were at 180 days. Endoprotease and exoprotease activities of anchovy sauces added with 20$\%$ and 30$\%$ of shrimp byproducts tend to be higher than those of control (0$\%$) and 10$\%$ addition. Proteolytic activities of sauces at pH 9 were about 2 times higher than those at pH 6. Amidolytic activities for LeuPNA decreased remarkably during fermentation, and control (0$\%$) almost lost their activity at 180 days, while additional sauces were relatively stable. These suggest that alkaline pretense of anchovy and shrimp byproducts as a endoprotease mainly contributed to the fermentation of salt-fermented sauces. The protein molecular weight distribution of sauces indicated 2 groups of peaks (peak 1,>70,000 da and peak 2, 3,000$\∼$29,000 da). As the fermentation proceeded, peak 1 tended to decrease in all of sauces, but peak 2 increased rapidly from 30 to 270 days. Optimum fermentation periods of control and 10$\%$ sauces were 270 days and those of 20$\%$ and 30$\%$ sauce were 180 days. The results suggest that shrimp byproduct can be used as accelerator of salt-fermented sauce.

Microbiological and Enzymological Studies on Takju Brewing (탁주(濁酒) 양조(釀造)에 관(關)한 미생물학적(微生物學的) 및 효소학적(酵素學的) 연구(硏究))

  • Kim, Chan-Jo
    • Applied Biological Chemistry
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    • v.10
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    • pp.69-100
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    • 1968
  • 1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.

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The Influence of Ammonium-Nitrogen on Anaerobic Microorganisms in Swine Wastewater by Batch-Fermentation. (혐기성 회분식 배양에서 양돈폐수의 NH$_4$-Nitrogen이 혐기성 미생물에 미치는 영향)

  • 김연옥
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.173-178
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    • 1998
  • This study presents the influence of ammonium-nitrogen on microorganisms in swine wastewater. For the anaerobic batch fermentation, two different methods were used. One is the dilution of wastewater with water. The other method is the elimination of ammonium-nitrogen from the wastewater. By addition of MgO into wastewater, non-soluble crystall was formed under alkaline condition as MgNH$_4$PO$_4$6$H_2O$ (MAP). The master culture was adapted in swine wastewater for more than 3 months, in water-dilution method, the dilution of wastewater with 25% water gave us the best result in efficiency of COD removal. Two hundred hours later MAP-treated wastewater showed the efficiency of the COD removal more than 80%. Under same condition obtained none MAP-treated wastewater about 50%. MAP treatment carried out the very effective anaerobic digestion with swine wastewater. The important result in this study is that the low ratio of C:N influenced on anaerobic microorganisms more than high concentration of ammonium nitrogen in swine wastewater. The struvite for the crystallforming has no toxic effect on methanogenic bacteria.

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Physicochemical Properties of a Biopolymer Flocculant Produced from Bacillus subtilis PUL-A (Bacillus subtilis PUL-A로부터 생산된 Biopolymer 응집제의 물리화학적 특성)

  • Ryu, Mi-Jin;Jang, Eun-Kyung;Lee, Sam-Pin
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.203-209
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    • 2007
  • Soybean milk cake (SMC) was used for the solid-state fermentation by Bacillus subtilis PUL-A isolated from soybean milk cake. In the presence of 5% glutamate the maximum production of biopolymer (59.9 g/kg) was performed by fermentation at $42^{\circ}C$ for 24 hr. The recovered biopolymer was consisted of 87% $\gamma$-polyglutamic acid with molecular weight of $1.3{\times}10^6$ dalton and other biopolymer. The biopolymer solution showed the great decrease in consistency below pH 6.0, regardless of the molecular weight of PGA. Biopolymer solution has a typical pseudoplastic flow behavior and yield stress. The consistency of biopolymer solution was greatly decreased by increasing heating time and temperature in acidic condition compared to the alkaline condition. In kaolin clay suspension, the flocculating activity of biopolymer was the highest value with 15 mg/L biopolymer and 4.5 mM $CaCl_2$, but decreased greatly with $FeCl_3$. The flocculating activity of biopolymer was maximum at pH5, but decreased drastically by heating at $60{\sim}100^{\circ}C$. In particular, biopolymer with native PGA showed the efficient flocculating activity compared to that of modified biopolymer containing low molecular weight of PGA.

Functional Properties of Soybean Curd Residue Fermented by Bacillus sp. (Bacillus firmus NA-1 균주와 Bacillus subtilis G7-D 균주를 이용한 발효비지의 기능성)

  • Oh, Soo-Myung;Kim, Chan-Shick;Lee, Sam-Pin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.115-120
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    • 2006
  • To convert the soybean curd residue (SCR) to functional food ingredient, alkaline fermentation of SCR was performed by Bacillus firmus NA-1 and Bacillus subtilis GT-D for 22 hr at $42^{\circ}C$. The micronized full-fat soy flour (MFS) was fortified to reduce the moisture content as well as to supply protein source. The mucilage and flavor productions in the fermented SCR were enhanced by the fortification of $20\%$ MFS. The peptide production from the SCR fermented with B. subtilis GT-D substantially increased when judged by the detectable amount of tyrosine $(480\;mg\%)$. The production of fibrinolytic enzyme was increased by the fermentation for 22 hr, indicating the relative activity of $62\%$ (B. firmus NA-1) and $47\%$ (B. subtilis GT-D), respectively. The SCR fermented by B. firmus NA-1 and B. subtilis GT-D indicated the consistency of $1.95\;Pa{\cdot}s^n\;and\;0.21\;Pa{\cdot}s^n$, respectively. After freeze-drying, the protease activity (615 unit/g) and a-amylase activity (180 unit/g) were obtained from SCR fermented by Bacillus firmus NA-1 and Bacillus subtilis GT-D, respectively.

Nutrient Recovery from Sludge Fermentation Effluent in Upflow Phosphate Crystallization Process (상향류 인 결정화공정을 이용한 슬러지 발효 유출수로 부터의 영양소 회수)

  • Ahn, Young-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.8
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    • pp.866-871
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    • 2006
  • The nutrient recovery in phosphate crystallization process was investigated by using laboratory scale uptlow reactors, adopting sequencing batch type configuration. The industrial waste lime was used as potential cation source with magnesium salt($MgCl_2$) as control. The research was focused on its successful application in a novel integrated sludge treatment process, which is comprised of a high performance fermenter followed by a crystallization reactor. In the struvite precipitation test using synthetic wastewater first, which has the similar characteristics with the real fermentation effluent, the considerable nutrient removal(about 60%) in both ammonia and phosphate was observed within $0.5{\sim}1$ hr of retention time. The results also revealed that a minor amount(<5%) of ammonia stripping naturally occurred due to the alkaline(pH 9) characteristic in feed substrate. Stripping of $CO_2$ by air did not increase the struvite precipitation rate but it led to increased ammonia removal. In the second experiment using the fermentation effluent, the optimal dosage of magnesium salt for struvite precipitation was 0.86 g Mg $g^{-1}$ P, similar to the mass ratio of the struvite. The optimal dosage of waste lime was 0.3 g $L^{-1}$, resulting in 80% of $NH_4-N$ and 41% of $PO_4-P$ removal, at about 3 hrs of retention time. In the microscopic analysis, amorphous crystals were mainly observed in the settled solids with waste lime but prism-like crystals were observed with magnesium salt. Based on mass balance analysis for an integrated sludge treatment process(fermenter followed by crystallization reactor) for full-scale application(treatment capacity Q=158,880 $m^3\;d^{-1}$), nutrient recycle loading from the crystallization reactor effluent to the main liquid stream would be significantly reduced(0.13 g N and 0.19 g P per $m^3$ of wastewater, respectively). The results of the experiment reveal therefore that the reuse of waste lime, already an industrial waste, in a nutrient recovery system has various advantages such as higher economical benefits and sustainable treatment of the industrial waste.

Development of Functional Vinegar by Using Cucumbers (오이를 이용한 기능성 식초 음료 개발)

  • Hong, Sung-Min;Moon, Hyun-Sil;Lee, Ju-Hye;Lee, Hae-In;Jeong, Ji-Hye;Lee, Mi-Kyung;Seo, Kwon-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.7
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    • pp.927-935
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    • 2012
  • This study was performed to develop functional vinegar by using cucumbers through two stages of fermentation. The alcohol content was maximized (7.8%) after 6-days of alcohol fermentation at $25^{\circ}C$ by adjusting the initial sugar concentration to $15^{\circ}Brix$, and vinegar with an acidity of 5.8% was obtained after 12-days of acetic acid fermentation at $30^{\circ}C$. The major sugars in the produced vinegar were glucose and fructose, which were present in concentrations of 3,067.26 and 395.73 mg%, respectively. The major organic acids were acetic acid and succinic acid, which were present in concentrations of 4,410.5 and 841.11 mg%, respectively. The total free amino acid content of the cucumber vinegar was 181.45 ${\mu}g/mL$ and citrulline, valine, aspartic acid, asparagine, and ornithine were the major amino acids. The inorganic components included various alkaline elements, such as K, Ca, and Mg. In addition, experimental methods to assess the DPPH and $ABTS^+$ radical-scavenging ability, reducing power, and ${\beta}$-carotene bleaching activity showed that the cucumber vinegar had strong antioxidant properties. The total polyphenol content, which are the major components responsible for the antioxidant activities of the cucumber vinegar, was 40.14 mg/100 mL. The cucumber vinegar showed significantly higher hepatic aldehyde dehydrogenase activity when compared to the alcoholic control (negative) and the marketing drink (positive), resulting in decreased plasma acetaldehyde concentrations in rats. These results demonstrate that cucumber vinegar possesses antioxidant properties and holds great promise for use in preventing hangovers.

Hepatoprotective and Curative Properties of Kombucha Tea Against Carbon Tetrachloride-Induced Toxicity

  • Murugesan, G.S.;Sathishkumar, M.;Jayabalan, R.;Binupriya, A.R.;Swaminathan, K.;Yun, S.E.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.397-402
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    • 2009
  • Kombucha tea(KT) is sugared black tea fermented with a symbiotic culture of acetic acid bacteria and yeasts, which is said to be tea fungus. KT is claimed to have various beneficial effects on human health, but there is very little scientific evidence available in the literature. In the present study, KT along with black tea(BT) and black tea manufactured with tea fungus enzymes(enzyme-processed tea, ET) were evaluated for hepatoprotective and curative properties against $CCl_4$-induced toxicity, using male albino rats as an experimental model by analyzing aspartate transaminase, alanine transaminase, and alkaline phosphatase in plasma and malondialdehyde content in plasma and liver tissues. Histopathological analysis of liver tissue was also included. Results showed that BT, ET, and KT have the potential to revert the $CCl_4$-induced hepatotoxicity. Among the three types of teas tried, KT was found to be more efficient than BT and ET. Antioxidant molecules produced during the fermentation period could be the reason for the efficient hepatoprotective and curative properties of KT against $CCl_4$-induced hepatotoxicity.