• Title/Summary/Keyword: Enzymatic Saccharification

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Industrial applications and characteristics of lignocellulolytic enzymes in Basidiomycetous fungi (담자균류 목질섬유소 분해효소의 특성과 산업적 이용)

  • Lim, Sun-Hwa;Kang, Hee-Wan
    • Journal of Mushroom
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    • v.14 no.2
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    • pp.51-58
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    • 2016
  • Basidiomycetous fungi are one of the most potent biodegraders because many of its species grow on dead wood or litter, in environments rich in lignocellulose. For the degradation of lignocellulose, basidiomycetes utilize their lignocellulytic enzymes, which typically include laccase (EC 1.10.3.2), lignin peroxidase (EC 1.11.1.14), xylanase (EC 3.2.1.8), and cellulase (EC 3.2.1.4). In recent years, the practical applications of basidiomycetes have ranged from the textile to the pulp and paper industries, and from food applications to bioremediation processes and industrial enzymatic saccharification of biomass. Recently, spent mushroom substrates of edible mushrooms have been used as sources of bulk enzymes to decolorize synthetic dyes in textile wastewater. In this review, the occurrence, mode of action, general properties, and production of lignocellulytic enzymes from mushroom species will be discussed. We will also discuss the potential applications of these enzymes.

Towards a Miniaturized Culture Screening for Cellulolytic Fungi and Their Agricultural Lignocellulosic Degradation

  • Arnthong, Jantima;Siamphan, Chatuphon;Chuaseeharonnachai, Charuwan;Boonyuen, Nattawut;Suwannarangsee, Surisa
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1670-1679
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    • 2020
  • The substantial use of fungal enzymes to degrade lignocellulosic plant biomass has widely been attributed to the extensive requirement of powerful enzyme-producing fungal strains. In this study, a two-step screening procedure for finding cellulolytic fungi, involving a miniaturized culture method with shake-flask fermentation, was proposed and demonstrated. We isolated 297 fungal strains from several cellulose-containing samples found in two different locations in Thailand. By using this screening strategy, we then selected 9 fungal strains based on their potential for cellulase production. Through sequence-based identification of these fungal isolates, 4 species in 4 genera were identified: Aspergillus terreus (3 strains: AG466, AG438 and AG499), Penicillium oxalicum (4 strains: AG452, AG496, AG498 and AG559), Talaromyces siamensis (1 strain: AG548) and Trichoderma afroharzianum (1 strain: AG500). After examining their lignocellulose degradation capacity, our data showed that P. oxalicum AG452 exhibited the highest glucose yield after saccharification of pretreated sugarcane trash, cassava pulp and coffee silverskin. In addition, Ta. siamensis AG548 produced the highest glucose yield after hydrolysis of pretreated sugarcane bagasse. Our study demonstrated that the proposed two-step screening strategy can be further applied for discovering potential cellulolytic fungi isolated from various environmental samples. Meanwhile, the fungal strains isolated in this study will prove useful in the bioconversion of agricultural lignocellulosic residues into valuable biotechnological products.

Enhancement of Saccharification Yield of Ulva pertusa Kjellman by High Pressure Homogenization Process for Bioethanol Production (구멍갈파래의 고압 균질 전처리 공정을 통한 바이오에탄올 생산용 당화수율 증진)

  • Choi, Woon-Yong;Lee, Choon-Geun;Ahn, Ju-Hee;Seo, Yong-Chang;Lee, Sang-Eun;Jung, Kyung-Hwan;Kang, Do-Hyung;Cho, Jeong-Sub;Choi, Geun-Pyo;Lee, Hyeon-Yong
    • KSBB Journal
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    • v.26 no.5
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    • pp.400-406
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    • 2011
  • This study was investigated to improve the saccharification yield of Ulva pertusa Kjellman by the high pressure homogenization process. It was found that the high pressure homogenization pretreatment effectively destructed the cell wall structures only by using water. The high pressure homogenization process was operated under various conditions such as 10000, 20000 or 30000 psi with different recycling numbers. The optimal condition was determined as 30000 psi and 2 pass of recycling numbers and the sugar conversion yields were 16.02 (%, w/w) of glucose and 14.70 (%,w/w) of xylose, respectively. In the case of enzymatic treating the hydrolyzates with 5 FPU/glucan of celullase and 100 units/mL of amyloglucosidase, 65.8% of carbohydrates was converted into glucose. Using the hydrolysates of Ulva pertusa Kjellman, 48.7% of ethanol was obtained in the culture S.cerevisiae. These results showed that the high pressure homogenization process could efficiently hydrolyze the marine resource by using only water for bioethanol production.

Effect of chemical input during wet air oxidation pretreatment of rice straw in reducing biomass recalcitrance and enhancing cellulose accessibility

  • Morone, Amruta;Chakrabarti, Tapan;Pandey, R.A.
    • Korean Journal of Chemical Engineering
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    • v.35 no.12
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    • pp.2403-2412
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    • 2018
  • The present study was aimed at evaluating the effect of variable sodium carbonate ($Na_2CO_3$) loading during wet air oxidation (WAO) pretreatment of rice straw in reducing biomass recalcitrance. The research study was intended to increase the cellulose recovery, hemicellulose solubilization, lignin removal in the solid fraction and limiting the generation of inhibitors in the liquid fraction while reducing the chemical input. The operating condition of $169^{\circ}C$, 4 bar, 18 min and 6.5 g/L $Na_2CO_3$ loading resulted in maximum cellulose recovery of 82.07% and hemicellulose solubilization and lignin removal of 85.43% and 65.42%, respectively, with a total phenolic content of 0.36 g/L in the liquid fraction. The crystallinity index increased from 47.69 to 51.25 along with enzymatic digestibility with an increase in $Na_2CO_3$ loading from 0 to 6.5 g/L as a result of removal of barriers for saccharification via effective cleavage of ether and ester bonds cross-linking the carbohydrates and lignin as indicated by FT-IR spectroscopy. A further increase in the $Na_2CO_3$ loading to 9.5 g/L did not significantly increase the sugar release. Thus, it was concluded that 6.5 g/L $Na_2CO_3$ during WAO is sufficient to increase the delignification and deacetylation, leading to significant changes in apparent cellulose crystallinity inter alia improvement in cellulose accessibility and digestibility of rice straw.

Characterization of Pretreatment for Barley straw by Alkaline Solutions (염기 용매를 이용한 보릿짚의 전처리 특성)

  • Kim, Kyoung-Seob;Kim, Jun Seok
    • Korean Chemical Engineering Research
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    • v.50 no.1
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    • pp.18-24
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    • 2012
  • Lignocellulose 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 residual biomass, e.q., Barley straw, because of their several superior aspects as Agriculture residual biomass; low lignin, high contents of carbohydrates. Barley straw consists of 39.78% cellulose (glucose), 22.56% hemicelluloses and 19.27% lignin. Pretreatment of barley straw using NaOH pretreatment solutions concentration with 2%, temperature $85^{\circ}C$ and reaction times 1 hr were investigates. $NH_4OH$ pretreatment condition was solutions concentration with 15%, temperature $60^{\circ}C$, and reaction times 24hr were investigates. Furthermore, enzymatic saccharification using cellulose at $50^{\circ}C$, pH 4.8, 180 rpm for conversion of cellulose contained in barley straw to monomeric sugar. The pretreatment of barley straw using NaOH and $NH_4OH$ can significantly improve enzymatic saccharification of barley straw by extract more lignin and increasing its accessibility to hydrolytic enzymes. The result showed NaOH pretreatment extracted yield of lignin was 24.15%. $NH_4OH$ pretreatment extracted yield of lignin was 29.09%. Shaccharification of barley straw pretreatment by NaOH for 72hr and pH 4.8 result in maximum glucose concentration 15.39g/L (58.40%) and by $NH_4OH$ for 72hr and pH 4.8 result in maximum glucose concentration 16.01g/L (64.78%).

Study on the pretreatment of rice hull to enhance enzymatic saccharification (왕겨의 효소 당화 증진을 위한 전처리 방법 연구)

  • Bark, Surn-Teh;Koo, Bon-Cheol;Choi, Yong-Hwan;Moon, Youn-Ho;Cha, Young-Lok;Kim, Jung Kon;An, Gi Hong;Park, Kwang-Geun;Park, Don-Hee
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.11a
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    • pp.112.1-112.1
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    • 2011
  • 쌀을 주식으로 하는 우리나라의 여건상 연간 추정치로 싸라기 약 12만톤, 미강 약 49만톤, 왕겨 약 79만톤의 벼 도정 부산물이 발생하고 있다. 본 연구에서는 벼 도정 부산물 중 비식량 자원인 왕겨를 대상으로 고효율 효소 당화를 위한 바이오매스 전처리 방법을 탐색하였다. 왕겨 원시료의 초기 조성은 셀룰로스 34.5%, 헤미셀룰로스 20.5%, 리그닌 25.3%, 회분 14.6%로 나타났는데, 억새 등 초본계 바이오매스와 비교하여 특이하게 높은 성분은 회분으로 이는 벼에 대한 규산질 비료의 시용에서 기인한 것이다. 바이오매스 전처리에 많이 사용되는 암모니아, 희황산 용매와 규산염에 침식성을 가지는 가성소다 용매를 이용하여 각 용매별 단독 및 알칼리-산 복합 처리 하였을 때 효소 가수분해 효율, 고상시료 성분변화 등을 상호 비교하였다. 예비실험을 통하여 암모니아 처리조건은 15%(w/w) $150^{\circ}C$ 20분, 가성소다 처리조건은 1.5%(w/w) $150^{\circ}C$ 20분, 희황산 처리조건은 1.0%(w/w) $150^{\circ}C$ 10분으로 설정하였다. 암모니아 단독, 희황산 단독, 암모니아-희황산 복합 처리 시료의 효소 가수분해 효율은 각각 37.8%, 39.1%, 42.8%로 약 40%선에서 큰 차이가 없었다. 반면 가성소다 단독, 가성소다-희황산 복합 처리시료의 효소 가수분해 효율은 각각 62.7%, 82.8%로 나타나 앞선 3가지 처리방법 대비 50%, 100%에 가까운 효소 가수분해 효율 향상을 보였다. 이 때 전처리 고상시료의 성분 변화를 살펴보면 회분 함량에서 큰 차이를 보였는데 암모니아 단독, 가성소다 단독, 희황산 단독, 암모니아-희황산 복합, 가성소다-희황산 복합 처리에서 각각 47.8%, 77.1%, 43.5%, 55.8%, 94.7%의 회분 성분 기각률(rejection rate)을 나타냈다. 이는 왕겨 효소 가수분해 효율의 최대 저해요인이 회분임을 추정할 수 있다. 왕겨 전처리 알칼리 용매는 암모니아보다 가성소다가 더 효과적이었고 희황산 복합 처리시 그 효과가 크게 상승하였다. 따라서 규산염(회분) 함량이 높은 바이오매스는 고온 고압 조건에서 가성소다 용액으로 처리한 후 그 고상분을 희황산 용액으로 복합 처리하는 시스템이 효소 당화 증진에 매우 유리함을 확인하였다.

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Application of β-1,3-Glucanase from Pyrococcus furiosus for Ethanol Production using Laminarin (Pyrococcus furiosus의 β-1,3-glucanase를 처리한 laminarin 분해 산물을 이용한 바이오 에탄올의 생산)

  • Kim, Dong-Gyun;Kim, Eun-Young;Kim, Yu-Ri;Kim, Joong-Kyun;Lee, Han-Seung;Kong, In-Soo
    • Journal of Life Science
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    • v.21 no.1
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    • pp.68-73
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    • 2011
  • $\beta$-1,3-glucanase from Pyrococcus furiosus was applied for the saccharification of laminarin, which is a major oligo-saccharide component of brown algae, and the reaction mixture produced from laminarin was utilized as a substrate for alcohol fermentation using yeast. To prepare the recombinant $\beta$-1,3-glucanase, a $\beta$-1,3-glucanase gene was overexpressed in Escherichia coli and purified. Laminarin was degraded to an oligo- and mono-saccharide, such as glucose, after reaction with the purified recombinant $\beta$-1,3-glucanase, and the products after enzymatic treatment were confirmed by TLC and HPLC analysis. Decomposed laminarin after enzyme reaction was only added to the medium as a C-source for yeast alcohol production reaction. 0.3% alcohol production was detected from the cultured broth by gas chromatography after 48 hr of incubation. Further evaluation for optimal conditions of saccharification and alcohol fermentation can be suggested, as well as the possibility of using this enzymatic method to produce ethanol using laminarin.

A Study on the Method of Manufacturing Lactic Acid from Seaweed Biomass (해조류 바이오매스로부터 Lactic acid를 제조하는 방법에 관한 연구)

  • Lee, Hakrae;Ko, Euisuk;Shim, Woncheol;Kim, Jongseo;Kim, Jaineung
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.28 no.1
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    • pp.1-8
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    • 2022
  • With the spread of COVID-19 worldwide, non-face-to-face services have grown rapidly, but at the same time, the problem of plastic waste is getting worse. Accordingly, eco-friendly policies such as carbon neutrality and sustainable circular economy are being promoted worldwide. Due to the high demand for eco-friendly products, the packaging industry is trying to develop eco-friendly packaging materials using PLA and PBAT and create new business models. On the other hand, Ulva australis occurs in large quantities in the southern seas of Korea and off the coast of Jeju Island, causing marine environmental problems. In this study, lactic acid was produced through dilute acid pretreatment, enzymatic saccharification, and fermentation processes to utilize Ulva australis as a new alternative energy raw material. In general, seaweeds vary in carbohydrate content and sugar composition depending on the species, harvest location, and time. Seaweed is mainly composed of polysaccharides such as cellulose, alginate, mannan, and xylan, but does not contain lignin. It is difficult to expect high extraction yield of the complex polysaccharide constituting Ulva australis with only one process. However, the fusion process of dilute acid and enzymatic saccharification presented in this study can extract most of the sugars contained in Ulva australis. Therefore, the fusion process is considered to be able to expect high lactic acid production yield when a commercial-scale production process is established.

A Study on the Method of Manufacturing Lactic Acid from Ginkgo Biloba Leaf Extraction Byproducts (은행잎 추출부산물로부터의 Lactic acid 제조법에 관한 연구)

  • Euisuk Ko;Hakrae Lee;Woncheol Shim;Soohyeon Lee;Sunjin Kim;Jaineung Kim
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.29 no.2
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    • pp.95-102
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    • 2023
  • Despite the easing of social distancing, demand for non-face-to-face services continues to rise. Recently, the EU is pursuing a comprehensive plastic use reduction by expanding the scope of plastic use regulations for packaging plastics according to the New Cyclical Economy Action Plan(NCEAP). In response to this trend, the packaging industry is moving away from conventional non-degradable/petroleum-based plastics and conducting research on packaging materials using biodegradable plastics such as PLA(Poly Lactic Acid), PBAT(Poly Butylene Adipate-co-butylene Terephthalate). On the other hand, ginkgo leaves occur in large quantities in Korea and act as a cause of slip accidents and flooding. In this study, a method to utilize ginkgo biloba leaf as a new alternative biomass resource was proposed by producing lactic acid through pretreatment, enzymatic saccharification, and fermentation processes. For the efficiency of lactic acid production, a comparative analysis of lignin content from before and after browning was performed. In addition, the degree of glucan extraction was evaluated by applying a pretreatment method using three catalysts: hot water, sulfuric acid, and sodium hydroxide. It is difficult to expect high production of lactic acid with single process. Therefore, an integrated process operation using both the pretreated hydrolyzate and the residual solid enzymatic saccharification solution must necessarily be applied.

Bioethanol Production from Seaweed Kappaphycus alvarezii by Simultaneous Saccharification and Fermentation (홍조류(Kappaphycus alvarezii)의 동시 당화 발효를 이용한 바이오에탄올의 생산)

  • Ra, Chae Hun;Kim, Sung-Koo
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.145-149
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    • 2016
  • Thermal acid hydrolysis pretreatment of Kappaphycus alvarezii was carried out with 12% (w/v) seaweed slurry and 180 mM H2SO4 at 140°C for 5 min. Utility of the thermotolerant yeast Kluyveromyces marxianus KCTC7150 was evaluated with respect to cell growth and ethanol fermentation at 40°C was close to optimal for enzymatic hydrolysis. This could lead to the integration of both the saccharification and fermentation processes. The levels of ethanol production by simultaneous saccharification and fermentation (SSF) with non-adapted and adapted K. marxianus KCTC7150 were 9.1 g/l with an ethanol yield (YEtOH) of 0.24 and 10.2 g/l with an ethanol yield (YEtOH) of 0.27 at 156 h, respectively. The two-phase SSF process was employed in this study to improve the efficiency of ethanol fermentation. Adapted K. marxianus KCTC7150 using the two-phase SSF process produced 13.5 g/l with an ethanol yield (YEtOH) of 0.35 at 96 h. Development of the two-phase SSF process could enhance the overall ethanol fermentation yields of the seaweed K. alvarezii.