• Title/Summary/Keyword: Enzymatic process

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Optimization of Combined Process of Enzymatic Hydrolysis and Solvent Extraction for Production of Lycopene from Elaeagnus umbellata (보리수 나무 열매로부터 라이코펜 생산을 위한 효소 분해 및 유기용매 추출 복합 공정의 최적화)

  • Oh, Yun Hye;Lee, Ju Mi;Chae, Hee Jeong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.4
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    • pp.293-299
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    • 2020
  • This study was undertaken to optimize combining the processes of enzymatic hydrolysis and extraction for lycopene production from autumn olive berry. The autumn olive berry was pulverized and suspended in water, followed by treatment with various hydrolytic enzymes including Ceremix, Celluclast, AMG, Viscozyme, Pectinex, Promozyme, Ultraflo and Tunicase. Reaction solutions were subjected to extraction by applying different organic solvents including acetone, ethyl acetate, hexane and chloroform. Highest yields of lycopene extraction were obtained with the Ceremix (hydrolysis enzyme) and chloroform (extraction solvent) combination. Subsequently, using this ideal combination, enzymatic hydrolysis conditions, including enzyme concentration, pH and temperature, were statistically optimized to 0.58%, 5.5 and 54.4℃, respectively, by applying the response surface method. The lycopene extraction yield increased 2.3-fold (22.6 mg/100g) by using the selected combined process. We propose that these results could be used for the future development of bioactive materials required for bio-health care products.

Study on Affecting Variables Appearing through Chemical Pretreatments of Poplar Wood (Populus euramericana) to Enzymatic Hydrolysis (이태리 포플러의 화학적 전처리 공정을 통한 효소가수분해 영향 인자 분석)

  • Koo, Bon-Wook;Park, Nahyun;Yeo, Hwanmyeong;Kim, Hoon;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.3
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    • pp.255-264
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    • 2009
  • To evaluate the effects of chemical pretreatments of lignocellulosic biomass on enzymatic hydrolysis process, Populus euramericana was pretreated for 1 hr with 1% sulfuric acid ($H_2SO_4$) at $150^{\circ}C$ and 1% sodium hydroxide (NaOH) at $160^{\circ}C$, respectively. Before the enzymatic hydrolysis, each pretreated sample was subjected to drying process and thus finally divided into four subgroups; dried or non-dried acid pretreated samples and dried or non-dried alkali pretreated samples and chemical and physical properties of them were analyzed. Biomass degradation by acid pretreatment was determined to 6% higher compared to alkali pretreatment. By the action of acid ca. 24.5% of biomass was dissolved into solution, while alkali degraded ca. 18.6% of biomass. However, reverse results were observed in delignification rates, in which alkali pretreatment released 2% more lignin fragment from biomass to the solution than acid pretreatment. Unexpectedly, samples after both pretreatments were determined to somewhat higher crystallinity than untreated samples. This result may be explained by selective disrupture of amorphous region in cellulose during pretreatments, thus the cellulose crystallinity seems to be accumulated in the pretreated samples. SEM images revealed that pretreated samples showed relative rough and partly cracked surfaces due to the decomposition of components, but the image of acid pretreated samples which were dried was similar to that of the control. In pore size distribution, dried acid pretreated samples were similar to the control, while that in alkali pretreated samples was gradually increased as pore diameter increased. The pore volume which increased by acid pretreatment rapidly decreased by drying process. Alkali pretreatment was much more effective on enzymatic digestibility than acid pretreatment. The sample after alkali pretreatment was enzymatically hydrolyzed up to 45.8%, while only 26.9% of acid pretreated sample was digested at the same condition. The high digestibility of the sample was also influenced to the yields of monomeric sugars during enzymatic hydrolysis. In addition, drying process of pretreated samples affected detrimentally not only to digestibility but also to the yields of monomeric sugars.

Process Development for the Recovery of Sialic Acid Fraction by Enzymatic Hydrolysis of Egg Yolk Protein (난황 단백질의 효소 가수분해에 의한 sialic acid의 회수 공정 개발)

  • Kang Byung Chul;Lee Kwang Hyun
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.9-14
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    • 2005
  • Batch enzymatic hydrolysis of egg yolk protein by protease was carried out at laboratory scale coupled to an ultrafiltration module. Effect of ethanol concentrations on the performance of enzymatic hydrolysis was studied to determine the optimum condition of recovery of hydrolysate. The enzymatic hydrolysis was conducted stepwise with following conditions, $50^{\circ}C$, pH 10.0 and pH 6.5. Ethanol concentration was changed from 10 to $40\%$ (w/w). As ethanol concentration was increased, the recovery yield of total solid and protein in enzymatic hydrolysate was also increased. The content of sialic acid and protein in hydrolysate was independent of ethanol concentration. We also investigated the effect of ethanol concentration on the performance of ultrafiltration. As the concentration of ethanol in yolk protein was increased, the recovery yield of product was increased. Ultra­filtration of egg yolk protein hydrolysate was conducted to increase the content of sialic acid. Four ultrafiltation modules were used in this study, and we evaluated the performance of the UF modules. When Amicon module was used, the recovery percentage of total solid in retentate was $6.0\%$, which is the highest among the modules used. In spite of the difference in the recovery yield of total solid, the purity of sialic acid in retentate was about $2.0\%$, which was 5 times higher than that in feed. It was concluded that the recovery yield and the purity of sialic acid did not correlate with the types of modules and the size of MWCO.

Cybernetic Modeling of Simultaneous Saccharification and Fermentation for Ethanol Production from Steam-Exploded Wood with Brettanomyces custersii

  • Shin Dong-Gyun;Yoo Ah-Rim;Kim Seung-Wook;Yang Dae-Ryook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1355-1361
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    • 2006
  • The simultaneous saccharification and fermentation (SSF) process consists of concurrent enzymatic saccharification and fermentation. In the present cybernetic model, the saccharification process, which is based on the modified Michaelis-Menten kinetics and enzyme inhibition kinetics, was combined with the fermentation process, which is based on the Monod equation. The cybernetic modeling approach postulates that cells adapt to utilize the limited resources available to them in an optimal way. The cybernetic modeling was suitable for describing sequential growth on multiple substrates by Brettanomyces custersii, which is a glucose- and cellobiose-fermenting yeast. The proposed model was able to elucidate the SSF process in a systematic manner, and the performance was verified by previously published data.

Pretreatment Characteristics of Ammonia Soaking Method for Cellulosic Biomass (암모니아 Soaking 방법을 이용한 섬유소계 바이오매스의 전처리 특성)

  • Park, Yong-Cheol;Kim, Jin-Woo;Kim, Jun-Seok
    • Korean Chemical Engineering Research
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    • v.49 no.3
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    • pp.292-296
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    • 2011
  • Liberation of fermentable sugars from lignocellulosic biomass is one of the key challenges in production of cellulosic ethanol. Aqueous ammonia cleaves ether and ester bonds in lignin carbohydrate complexes. It is an effective swelling reagent for lignocellulosic biomass. The aqueous ammonia pretreatment selectively reduces the lignin content of biomass. However, at high temperatures, this process solubilizes more than 50% of the hemicellulose in the biomass. Here we conducted a SAA(Soaking in Aqueous Ammonia) process by moderate reaction temperatures at atmospheric pressure using various lignocellulosicbiomass. The optimum condition of this process was 15 wt% of aqueous ammonia at 50 of reaction time during 72 hr. The delignification was up to 60% basis on initial biomass and the enzymatic digestibility was 60-90% for agricultural biomass, respectively.

Enzymatic Milling Process for Barley Flour Preparation (보리의 효소적(酵素的) 제분(製粉) 및 이용(利用)에 관한 연구)

  • Kwon, Tai-Wan;Ahn, Byung-Yoon;Choi, Weon-Sang;Cheigh, Hong-Sik
    • Korean Journal of Food Science and Technology
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    • v.18 no.3
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    • pp.197-203
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    • 1986
  • An enzymatic flour milling process for barley into three major fractions (barley flour, bran-crease-germ and water solubles) was studied. Carbohydrate and protein of barley endosperm could be efficiently solubilized by the digestion process of partially pearled barley with enzymes. Bran, crease and germ were removed from hydrolyzate by filtering through 30-40mesh sieves. And then filtered product was separated into fractions by sedimentation or centrifugation. The most effective digestion of the barley was obtained by the enzyme with higher activities of glucanase and protease under such conditions as barley-water ratio, 1:1.5(W/V) and temperature at $45^{\circ}C$. Total flour yield recovered was approximately 73-76% of the barley, and the portions recovered as bran-crease-germ and water solubles were about 3.6 and 15.8%, respectively.

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A Color-Reaction-Based Biochip Detection Assay for RIF and INH Resistance of Clinical Mycobacterial Specimens

  • Xue, Wenfei;Peng, Jingfu;Yu, Xiaoli;Zhang, Shulin;Zhou, Boping;Jiang, Danqing;Chen, Jianbo;Ding, Bingbing;Zhu, Bin;Li, Yao
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.180-189
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    • 2016
  • The widespread occurrence of drug-resistant Mycobacterium tuberculosis places importance on the detection of TB (tuberculosis) drug susceptibility. Conventional drug susceptibility testing (DST) is a lengthy process. We developed a rapid enzymatic color-reaction-based biochip assay. The process included asymmetric multiplex PCR/templex PCR, biochip hybridization, and an enzymatic color reaction, with specific software for data operating. Templex PCR (tem-PCR) was applied to avoid interference between different primers in conventional multiplex-PCR. We applied this assay to 276 clinical specimens (including 27 sputum, 4 alveolar lavage fluid, 2 pleural effusion, and 243 culture isolate specimens; 40 of the 276 were non-tuberculosis mycobacteria specimens and 236 were M. tuberculosis specimens). The testing process took 4.5 h. A sensitivity of 50 copies per PCR was achieved, while the sensitivity was 500 copies per PCR when tem-PCR was used. Allele sequences could be detected in mixed samples at a proportion of 10%. Detection results showed a concordance rate of 97.46% (230/236) in rifampicin resistance detection (sensitivity 95.40%, specificity 98.66%) and 96.19% (227/236) in isoniazid (sensitivity 93.59%, specificity 97.47%) detection with those of DST assay. Concordance rates of testing results for sputum, alveolar lavage fluid, and pleural effusion specimens were 100%. The assay provides a potential choice for TB diagnosis and treatment.

Enhancement of Enzymatic Hydrolysis of Lignocellulosic Biomass by Organosolv Pretreatment with Dilute Acid Solution (효소당화를 위한 목질계 바이오매스의 유기용매 침출 전처리 공정)

  • Kim, Jun Beom;Kim, Jun Seok
    • Korean Chemical Engineering Research
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    • v.54 no.6
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    • pp.806-811
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    • 2016
  • Organosolv pretreatment is the process to frationation of lignocellulosic feedstocks to enhancement of enzymatic hydrolysis. This process has advantages that organic solvents are always easy to recover by distillation and recycled for pretreatment. The chemical recovery in organosolv pretreatment can isolate lignin as a solid material and carbohydrates as fermentable sugars. For the economic considerations, using of low-molecular-weight alcohols such as ethanol and methanol have been favored. When acid catalysts are added in organic solvent, the rate of delignification could be increased. Mineral acids (hydrochloric acid, sulfuric acid, and phosphoric acid) are good catalysts to accelerate delignification and xylan degradation. In this study, the biomass was pretreated using 40~50 wt% ethanol at $170{\sim}180^{\circ}C$ during 20~60 min. As a results, the enzymatic digestibility of 2-stage pretreatment of rigida using 50 wt% ethanol at $180^{\circ}C$ was 40.6% but that of 1-stage pretreatment was 55.4% on same conditions, therefore it is shown that the pretreatment using mixture of the organosolv and catalyst was effective than using them separately.

Optimization of Ammonia Percolation Process for Ethanol Production from Miscanthus Sinensis (억새를 이용한 바이오 에탄올 생산을 위한 암모니아 침출 공정 최적화)

  • Kim, Kyoung-Seob;Kim, Jun Seok
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.704-711
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    • 2010
  • Lignocellulose ($2^{nd}$ generation) is difficult to hydrolyze due to the presence of lignin and the technology developed for cellulose fermentation to ethanol is not yet economically viable. However, recent advances in the extremely new field of biotechnology for the ethanol production are making it possible to use of agriculture residuals and nonedible crops biomass, e.q., rice straw and miscanthus sinensis, because of their several superior aspects as agriculture residual and nonedible crops biomass; low lignin, high contents of carbohydrates. In this article, as the basic study of AP(Ammonia Percolation), the properties and the optium conditions of process were established, and then the overall efficiency of AP was investigated. The important independent variables for AP process were selected as ammonia concentration, reaction temperature, and reaction time. The percolation condition for maximizing the content of cellulose, the enzymatic digestibility, and the lignin removal was optimized using RSM(Response Surface Methodology). The determined optimum condition is ammonia concentration; 11.27%, reaction temperature; $157.75^{\circ}C$, and reaction time; 10.01 min. The satisfying results were obtained under this optimized condition, that is, the results are as follows: cellulose content(relative); 39.98%, lignin content(relative); 8.01%, and enzymatic digestibility; 85.89%.

Biological Pretreatment of Softwood Pinus densiflora by Three White Rot Fungi

  • Lee, Jae-Won;Gwak, Ki-Seob;Park, Jun-Yeong;Park, Mi-Jin;Choi, Don-Ha;Kwon, Mi;Choi, In-Gyu
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.485-491
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    • 2007
  • The effects of biological pretreatment on the Japanese red pine Pinus densiflora, was evaluated after exposure to three white rot fungi Ceriporia lacerata, Stereum hirsutum, and Polyporus brumalis. Change in chemical composition, structural modification, and their susceptibility to enzymatic saccharification in the degraded wood were analyzed. Of the three white rot fungi tested, S. hirsutum selectively degraded the lignin of this sortwood rather than the holocellulose component. After eight weeks of pretreatment with S. hirsutum, total weight loss was 10.7%, while lignin loss was the highest at 14.52% among the tested samples. However, holocellulose loss was lower at 7.81 % compared to those of C. lacerata and P. brumalis. Extracelluar enzymes from S. hirsutum showed higher activity of ligninase and lower activity of cellulase than those from other white rot fungi. Thus, total weight loss and changes in chemical composition of the Japanese red pine was well correlated with the enzyme activities related with lignin- and cellulose degradation in these fungi. Based on the data obtained from analysis of physical characterization of degraded wood by X-ray Diffractometry (XRD) and pore size distribution, S. hirsutum was considered as an effective potential fungus for biological pretreatment. In particular, the increase of available pore size of over 120 nm in pretreated wood powder with S. hirsutum made enzymes accessible for further enzymatic saccharification. When Japanese red pine chips treated with S. hirsutum were enzymatically saccharified using commercial enzymes (Cellulclast 1.5 L and Novozyme 188), sugar yield was greatly increased (21.01 %) compared to non-pre treated control samples, indicating that white rot fungus S. hirsutum provides an effective process in increasing sugar yield from woody biomass.