• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2005.06a
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• pp.245-247
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• 2005

A cellulolytic fungus isolated from Agave plantation in northeastern Thailand was identified as Acrophialophora nainiana. The fungus was capable of growing at pH between 3 - 7 and 25 - 45 $^{\circ}C$, with the optimum conditions at pH 5.0 and 40 $^{\circ}C$. The wild isolate produced cellulases, comprising of exoglucanase (0.019 U/mg protein), endoglucanase (0.366 U/mg protein), and ${\beta}$-glucosidase (0.001 U/mg protein). Mutations with UV and NTG produced the UV 10-2 mutant with cellulases activities including exoglucanase (0.093 U/mg protein), endoglucanase (0.585 U/mg protein), and ${\beta}$-glucosidase (0.013 U/mg protein). Purification of the enzymes with ultrafiltration, ammonium sulfate precipitation, and ion-exchange chromatography yielded the maximal cellulase specific activities of 2.736 U/mg protein (exoglucanase), 0.235 U/mg protein (endoglucanase), and 0.008 U/mg protein (${\beta}$-glucosidase). The mutant's cellulases were the most active at pH 5.0 and 60 $^{\circ}C$. Ion-exchange chromatography revealed that A. nainiana UV 10-2 cellulases were comprised of two peaks with one peak showing the single endoglucanase activity while the other peak showed a mixture of the three enzyme activities. Production of A. nainiana UV 10-2 cellulases using banana leaf stalk as the sole carbon source gave comparable yields to that of the pure ${\alpha}$-cellulose. The enzymes were used in the simultaneous saccharification and fermentation (SSF) of plant residue (Coix aquatica) along with Kluveromyces marxianus to produce ethanol. Moreover, when the enzymes were used in the bioscouring process of fabric, the desiravle traits of textile processing including immediate water absorbency, increased in whiteness and reduction of yellowness of the treated fabric were observed.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.260-267
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• 1990

The mechanism of enzymatic hydrolysis of raw corn starch by the purified glucoamylase and a - amylase in an agitated bead reaction system was studied by investigating the changes of sugar profiles produced by each enzyme, the granular structure of raw corn starch, the amount of enzyme adsorption on residual starch, and the amylose content in residual raw starch. The sugar profiles produced by the action of exo-type glucoamylase or endo-type $\alpha$ -amylase in an agitated bead system were not recognizably differed with those produced in reaction system without bead. Without enzyme the intergenic microcrystalline structure of starch granule was not changed by the simple mechanical impact of solid media, but it was cleaved. However, starch granule was fragment into large number of small particles by the synergistic action of enzyme and attrition-milling media, identified to be the major saccharification enhancing mechanism along with the increased amount of enzyme adsorption. The amylose content decreased more readily in an agitated bead reaction system, especially by $\alpha$ -amylase.

• New & Renewable Energy
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• v.6 no.4
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• pp.6-14
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• 2010

Pretreatment of cellulosic biomass is necessary before enzymatic saccharification and fermentation. Extrusion is a well established process in food industries and it can be used as a physicochemical treatment method for cellulosic biomass. Aqueous ammonia soaking treatment at mild temperatures ranging from 60 to $80^{\circ}C$ for longer reaction times has been used to preserve most of the cellulose and hemicellulose in the biomass. The objective of this study was to evaluate the effect of extrusion treatment on aqueous ammonia soaking method. Extrusion was performed with miscanthus sample conditioned to 2mm of particle size and 20% of moisture content at $200^{\circ}C$ of barrel temperature and 175rpm of screw speed. And then aqueous ammonia soaking was performed with 15%(w/w) ammonia solution at $60^{\circ}C$ for 1, 2, 4, 8, 12 hours on the extruded and raw miscanthus samples respectively. In the combined extrusion-soaking treatment, most compositions removal occurred within 1~2 hours and on a basis of 1 hour soaking treatment values, cellulose was recovered about 85% and other compositions, including hemicellulose, are removed about 50% from extruded miscanthus sample. The combined extrusion-soaking treated and soaking only treated samples were subjected to enzymatic hydrolysis using cellulase and ${\beta}$-glucosidase. The enzymatic digestibility value of combined extrusion-2 hours soaking treated sample was comparable to 12 hours soaking only treated sample. It means that extrusion treatment can shorten the conventional long reaction time of aqueous ammonia soaking. The findings suggest that the combination of extrusion and soaking is a promising pretreatment method to solve both problems for no lignin removal of extrusion and long reaction time of aqueous ammonia soaking.

• Journal of the Korean Wood Science and Technology
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• v.14 no.3
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• pp.23-29
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• 1986

Alakline peroxide pretreatment for the delignification of poplar wood was performed. sinceit is a simple and efficent method for enhancing the enzymatic digestibility of wood residues. Approximately one-half of their lignin and most of the hemicellulose present in poplar wood were removed when the wood sawdust was reacted at 25$^{\circ}C$ for 100 hrs in an alkaline solution (pH 11.5) of 1% peroxide. The rate of decomposition as well as the saccharification efficiency were enhanced up to 350% and 260% respectively in comparision with those of the controll. This enhancement is comparable with that pretreated with 1% sodium hydroxide and 20% peracetic acid successively. The advantages of alkaline peroxide as delignifying agents against other chemicals were also discussed.

• 한국신재생에너지학회:학술대회논문집
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• 2011.11a
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• pp.114.2-114.2
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• 2011

Furfural is a versatile derivative. It can be utilized for a building-block of furfuryl alcohol production and a component of fuels or liquid alkanes. But in bio-process, furfural is a critical compound because it inhibits cell growth and metabolism. Furfural could be converted from xylose and usually produced from biomass in which hemicellulose is abundant. In this study, furfural production from miscanthus was performed and utilization of miscanthus residue was consequently conducted. At first, hydrolysis for investigation of miscanthus composition and furfural production was performed using sulfuric acid. Previously, we optimized dilute acid pretreatment condition for miscanthus pretreatment and the condition was found to be about 15 min of reaction time, 1.5% of acid concentration and about $140^{\circ}C$ of temperature and 60% (about 7 g/L) of xylose was solubilized from miscanthus. Using the xylose, furfural production was conducted as second step. Approximately $160{\sim}200^{\circ}C$ of temperature was accompanied with the hydrolysis for pyrolysis of biomass. When the investigated condition; $180^{\circ}C$ of temperature, 20 min of reaction time and 2% of acid concentration was operated for furfural production, furfural productivity was reached to be 77% of theoretical maximum. After reaction, residue of miscanthus was utilized as feedstock of ethanol fermentation. Residue was well washed using water and saccharified using hydrolysis enzymes. Hydrolysate (glucose) from saccharification was utilized for the carbon source of Saccharomyces cervisiae K35.

• 한국신재생에너지학회:학술대회논문집
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• 2011.11a
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• pp.114.1-114.1
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• 2011

Bioenergy production from lignocellulosic biomass and agriculture wastes have been attracted because of its sustainable and non-edible source. Especially, canola is considered as one of the best feedstock for renewable fuel production. Oil extracted canola and its agriculture residues are reuseable for bioethanol production. However, a pretreatment step is required before enzymatic hydrolysis to disrupt recalcitrant lignocellulosic matrix. To increase the sugar conversion, more efficient pretreatment process was necessary for removal of saccharification barriers such as lignin. Alkaline pretreatment makes the lignocellulose swollen through solvation and induces more porous structure for enzyme access. In our previous work, aqueous ammonia (1~20%) was utilized for alkaline reagent to increase the crystallinity of canola residues pretreatment. In this study, significant factors for efficient soaking in aqueous ammonia pretreatment on canola residues was optimized by using the response surface method (RSM). Based on the fundamental experiments, the real values of factors at the center (0) were determined as follows; $70^{\circ}C$ of temperature, 17.5% of ammonia concentration and 18 h of reaction time in the experiment design using central composition design (CCD). A statistical model predicted that the highest removal yield of lignin was 54% at the following optimized reaction conditions: $72.68^{\circ}C$ of temperature, 18.30% of ammonia concentration and 18.30 h of reaction time. Finally, maximum theoretical yields of soaking in aqueous ammonia pretreatment were 42.23% of glucose and 22.68% of xylose.

• Journal of Mushroom
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• v.9 no.2
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• pp.80-83
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• 2011

Sparassis crispa(Cauliflower mushroom) was an edible mushroom that shows remarkably high contents of ${\beta}$-glucan compared to other edible mushroom. In this study, we aimed to select suitable conditions and materials for mycelial growth and fruit body production. The longer saccharification time of barely result in higher sugar content for eight hours. The optimal sugar content of media for mycelial growth was showed at level of 6 $^{\circ}Brix$. Optimum temperature and pH for mycelial growth in liquid spawn were $25^{\circ}C$ and pH 5.0~6.0, respectively. In case of sawdust was used Larix kaempferi as main material, the fruit body yield and cultivation period of supplemented with 10% wheat flour and 20% corn flour were highest and fastest, respectively.

• Journal of Applied Biological Chemistry
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• v.60 no.4
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• pp.333-338
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• 2017

Cellulosic alcohol fermentation has recently gained more attention in the production of ethanol, butanol, and 2,3-butanediol. However, it was revealed that the process had several hurdles, such as, an expensive cost for biomass decomposition to yield fermentable sugars and a production of byproduct lignin. As an alternative for the process through biomass saccharification, the alcohol production through syngas from biomass has been studied. In this study, we reviewed acetogen and its central metabolic pathway, Wood-Ljungdahl route, capable of utilizing syngas. Furthermore, the metabolic engineering strategies of acetogen for bio-alcohol production from syngas was also reviewed with a brief perspective.

• Applied Biological Chemistry
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• v.3
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• pp.9-15
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• 1962

1) Among 8 strains of Rhizopus and 7 of Aspergillus species investigated for their producibility of saccharifying amylase, Rhizopus delemar ND 1 and Aspergillus usamii mut. shirousamii were selected as having high saccharifying activity. 2). Since Rh. delemar ND 1 shows high saccharifying activity and slight transglucosidase activity, it likely seems suitable for the production of starch-sugar by enzymic saccharification. Asp. usamii mut. shirousamii exerts low saccharifying but moderate transglucesidase activity and is considered to be usable in producing sugar-syrup with particular flavor and taste. 3). Using the saccharogenic enzyme from Rh. delemar ND 1, a prelimimary trial was done to obtain purified glucose powders.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.19-19
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• 2014

Pleurotus ostreatus, P. eryngii, and Flammulina velutipes are major edible mushrooms that account for over 89% of total mushroom production in Korea. Recently, Agrocybe cylindracea, Hypsizygus marmoreus, and Hericium erinaceu are increasingly being cultivated in mushroom farms. In Korea, the production of edible mushrooms was estimated to be 614,224 ton in 2013. Generally, about 5 kg of mushroom substrate is needed to produce 1 kg of mushroom, and consequently about 25 million tons of spent mushroom substrate (SMS) is produced each year in Korea. Because this massive amount of SMC is unsuitable for reuse in mushroom production, it is either used as garden fertilizer or deposited in landfills, which pollutes the environment. It is reasonably assumed that SMS includes different secondary metabolites and extracellular enzymes produced from mycelia on substrate. Three major groups of enzymes such as cellulases, xylanases, and lignin degrading enzymes are involved in breaking down mushroom substrates. Cellulase and xylanase have been used as the industrial enzymes involving the saccharification of biomass to produce biofuel. In addition, lignin degrading enzymes such as laccases have been used to decolorize the industrial synthetic dyes and remove environmental pollutions such as phenolic compounds. Basidiomycetes produce a large number of biologically active compounds that show antibacterial, antifungal, antiviral, cytotoxic or hallucinogenic activities. However, most previous researches have focused on therapeutics and less on the control of plant diseases. SMS can be considered as an easily available source of active compounds to protect plants from fungal and bacterial infections, helping alleviate the waste disposal problem in the mushroom industry and creating an environmentally friendly method to reduce plant pathogens. We describe extraction of lignocellulytic enzymes and antimicrobial substance from SMSs of different edible mushrooms and their potential applications.