• Journal of Appropriate Technology
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• v.5 no.2
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• pp.91-96
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• 2019

Microalgae do not require much land and make a higher efficient oil production. However, it costs still much higher than other biodiesel resources, such as crops. Sugars charge 80% of culture media when microalgae are massively cultured in the fermenter. This study aims to develop a cost-efficient process for sugar production from Chinese cabbage byproducts. Pre-treatment with 0.25% H₂SO₄ was most effective when chopped cabbage was incubated 50℃/130 rpm for 24 hours. To hydrolyze cabbage cellulose, we used cellulases secreted from Trichoderma. harzianum. T. harzianum was cultured at 28℃/pH 7/130 rpm for five days. Optimal enzymatic activity of cellulase was obtained by incubating at 0.24 FPU/ml/45℃/pH 5/130 rpm for three days. In comparison to other agricultural waste, such as rice straw, green tea leaves, and palm residue, Chinese cabbage produced the highest sugar yield. We found the optimal conditions to produce sugar from Chinese cabbage byproducts as a carbon source to culture biodiesel-producing microalgae. The efficient process developed in this study helps microalgae as a sustainable alternative energy source by cost-down.

• KSBB Journal
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• v.11 no.3
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• pp.317-322
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• 1996

Galactooligosaccharlde (GOS) is a kind of functional oligosaccharides that can be used as a food ingredient and a cosmetic additive. In this paper, characteristics of GOS synthesis by cellulase, using lactose as a substrate, were investigated. Penicillium funiculosum cellulose was found to be the most efficient for GOS production among six cellulose tested. The optimum pH and temperature for GOS production were 5.0 and $50^{\circ}C$, respectively. There was an optimum ratio of lactose concentration to enzyme loading; the value was 10 (w/w). The reaction pattern of P. funiculosum cellulase is consistent with that of microbial ${\beta}$-galactosidase which shows transgalactosylation activity. Amounts of GOS produced from 20% (w/v) lactose after 6 h incubation at $50^{\circ}C$, were 23% (w/w) based on total saccharide in the reaction medium. The GOS % increased with initial lactose concentration in the range of 5 to 20%. The products mainly consisted of a trisaccharide and tetrasaccharide from HPLC and TLC analysis. Among enzymes involved in transgalactosylation reaction, high molecular weight fractions over 50,000 Da, presumably ${\beta}$-glucosldase, were considered to be responsible for GOS production. Using this cellulose, a direct synthesis of galactosyl g1ucoside including GOS could be readily achieved with lactose as a galactosyl donor.

• Journal of Plant Biotechnology
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• v.34 no.2
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• pp.111-118
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• 2007

Global warming crisis due primarily to continued green house gas emission requires impending change to renewable alternative energy than continuously depending on exhausting fossil fuels. Bioenergy including biodiesel and bioethanol are considered good alternatives because of their renewable and sustainable nature. Bioethanol is currently being produced by using sucrose from sugar beet, grain starches or lignocellulosic biomass as sources of ethanol fermentation. However, grain production requires significant amount of fossil fuel inputs during agricultural practices, which means less competitive in reducing the level of green house gas emission. By contrast, cellulosic bioethanol can use naturally-growing, not-for-food biomass as a source of ethanol fermentation. In this respect, cellulosic ethanol than grain starch ethanol is considered a more appropriate as a alternative renewable energy. However, commercialization of cellulosic ethanol depends heavily on technology development. Processes such as securing enough biomass optimized for economic processing, pretreatment technology for better access of polymer-hydrolyzing enzymes, saccharification of recalcitrant lignocellulosic materials, and simultaneous fermentation of different sugars including 6-carbon glucose as well as 5-carbon xylose or arabinose waits for greater improvement in technologies. Although it seems to be a long way to go until commercialization, it should broadly benefit farmers with novel source of income, environment with greener and reduced level of global warming, and national economy with increased energy security. Mission-oriented strategies for cellulosic ethanol development participated by government funding agency and different disciplines of sciences and technologies should certainly open up a new era of renewable energy.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.336-343
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• 2016

A mannanase gene was cloned into Escherichia coli from Cellulosimicrobium sp. YB-43, which had been found to produce two kinds of mannanase, and sequenced completely. This mannanase gene, designated manB, consisted of 1,284 nucleotides encoding a polypeptide of 427 amino acid residues. Based on the deduced amino acid sequence, the ManB was identified to be a modular enzyme including two carbohydrate binding domains besides the catalytic domain, which was highly homologous to mannanases belonging to the glycosyl hydrolase family 5. The N-terminal amino acid sequence of ManB, purified from a cell-free extract of the recombinant E. coli carrying a Cellulosimicrobium sp. YB-43 manB gene, has been determined as QGASAASDG, which was correctly corresponding to signal peptide predicted by SignalP4.1 server for Gram-negative bacteria. The purified ManB had a pH optimum for its activity at pH 6.5~7.0 and a temperature optimum at $55^{\circ}C$. The enzyme was active on locust bean gum (LBG), konjac and guar gum, while it did not exhibit activity towards carboxymethylcellulose, xylan, starch, and para-nitrophenyl-${\beta}$-mannopyranoside. The activity of enzyme was inhibited very slightly by $Mg^{2+}$, $K^+$, and $Na^+$, and significantly inhibited by $Cu^{2+}$, $Zn^{2+}$, $Mn^{2+}$, and SDS. The enzyme could hydrolyze mannooligosaccharides larger than mannobiose, which was the most predominant product resulting from the ManB hydrolysis for mannooligosaccharides and LBG.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.329-336
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• 2021

Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 ℃. Its activity increased by more than six times in the presence of Co²⁺ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

• 한국신재생에너지학회:학술대회논문집
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• 2011.05a
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• pp.179.1-179.1
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• 2011

Pretreatment of cellulosic biomass is necessary before enzymatic saccharification and fermentation. The objective of this study was to evaluate the effect of combined aqueous ammonia-dilute sulfuric acid treatment on cellulosic biomass. Miscanthus was pretreated using aqueous ammonia and dilute sulfuric acid solution under high temperature and pressure conditions to be converted into bioethanol. Aqueous ammonia treatment was performed with 15 %(w/w) ammonia solution at $150^{\circ}C$ of reaction temperature and 20 minutes of reaction time. And then, dilute sulfuric acid treatment was performed with 1.0 %(w/w) sulfuric acid solution at $150^{\circ}C$ of reaction temperature and 10 minutes of reaction time. The compositional variations of this combined aqueous ammonia-dilute sulfuric acid treatment resulted in 68.0 % of cellulose recovery and 95.7 % of hemicellulose, 81.3 % of lignin, 89.1 % of ash removal respectively. The enzymatic digestibility of 90.5 % was recorded in the combined pretreated Miscanthus sample and it was 14.7 times higher than the untreated sample. The ethanol yield in the Simultaneous Saccharification and Fermentation was 90.4 % of maximum theoretical yield based on cellulose content of the combined pretreated sample and it was about 98 % compared to the ${\alpha}$-cellulose ethanol yield.

• 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.33 no.6 s.134
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• pp.79-86
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• 2005

Aspen wood (Populus tremuloides, L.) was biotreated with Ceriporiopsis subvermispora for 1, 2, 4, and 6 weeks to observe the physical/chemical modification of wood components. Milled wood lignins (MWLs) isolated from each decayed wood were analyzed by gel permeation chromatography (GPC) and nitrobenzene oxidation (NBO). As fungal treatment was progressed, lignin contents continuously decreased up to 20% after 6-week treatment. The lignin polymer could be fragmented to low-molecular phenolics, which make an enhancement of alkali solubility. Holocellulose contents were not affected severely during the period of fungal treatment, only reduction of 5~6% compared to the control. Xylose contents were decreased gradually from 23.4% to 18% after 6 weeks, whereas alpha-cellulose remained almost unchanged. Gel permeation chromatography (GPC) indicates that molecular weight of lignin undergoes a slight decrement for 4 weeks of fungal treatment. Nitrobenzene oxidation revealed that total yield of NBO products of lignins were lowered ca 20% after fungal treatment. Sum of syringaldehyde and syringic acid are remarkably decreased. However, increment of sum of vanillin and vanillic acid was surprisingly observed. These results work as indirect evidence that a specific lignolytic reaction, maybe selective demethoxylaytion of S-lignin, can occur during fungal treatment of aspen wood by C. subvermispora.

• Microbiology and Biotechnology Letters
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• v.45 no.2
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• pp.155-161
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• 2017

A gene coding for the xylanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned by PCR amplification and sequenced completely. This xylanase gene, designated xyn11B, consisted of 1,071 nucleotides encoding a polypeptide of 356 amino acid residues. Based on the deduced amino acid sequence, Xyn11B was identified to be a modular enzyme, including a single carbohydrate-binding module besides the catalytic domain, and was highly homologous to xylanases belonging to glycosyl hydrolase family 11. The SignalP4.1 server predicted a stretch of 26 residues in the N-terminus to be the signal peptide. Using DEAE-Sepharose and Phenyl-Sepharose column chromatography, Xyn11B was partially purified from the cell-free extract of recombinant Escherichia coli carrying a copy of the P. woosongensis xyn11B gene. The partially purified Xyn11B protein showed maximal activity at $50^{\circ}C$ and pH 6.5. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchwood xylan, whereas it did not exhibit activity towards carboxymethylcellulose, mannan, and para-nitrophenyl-${\beta}$-xylopyranoside. The activity of Xyn11B was slightly increased by $Ca^{2+}$ and $Mg^{2+}$, but was significantly inhibited by $Cu^{2+}$, $Ni^{2+}$, $Fe^{3+}$, and $Mn^{2+}$, and completely inhibited by SDS.