• Journal of Korean Society of Water and Wastewater
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• v.14 no.1
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• pp.117-126
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• 2000

Solubilization pretreatments were conducted to enhance the anaerobic digestion of the waste activated sludge. Four pretreatment techniques including heating, sonication freezing and thawing, and enzyme addition were employed to solubilize the waste activated sludge under various conditions. Thermal pretreatment by heating showed the highest efficiency compared with other methods, and freezing and thawing was confirmed as a feasible alternative of solubilization as well as the pretreatment of dewatering. There is a clear correlation between the solubilization efficiency of the waste activated sludge and the gas production. Batch digestion results showed the cumulative gas production as much as four times after thermal pretreatment as compared with that by the control sludge without pretreatment. As a result, hydrolysis or solubilization pretreatment might play a significant role in the high rate digestion of the waste activated sludge.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.7
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• pp.915-922
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• 2009

Ruminants possess the capacity to digest very large amounts of starch. However, in many cases diets approach 60% starch and even small inefficiencies present opportunities for energetic losses. Ruminal starch digestion is typically 75-80% of starch intake. On average, 35-60% of starch entering the small intestine is degraded. Of the fraction that escapes small-intestinal digestion, 35-50% is degraded in the large intestine. The low digestibility in the large intestine and the inability to reclaim microbial cells imposes a large toll on post-ruminal digestive efficiency. Therefore, digestibility in the small intestine must be optimized. The process of starch assimilation in the ruminant is complex and remains an avenue by which increases in production efficiency can be gained. A more thorough description of these processes is needed before we can accurately predict digestion occurring in the small intestine and formulate diets to optimize site of starch digestion.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.113-116
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• 1997

The hydrogel process is a different form of ultrafiltration and has been used to separate biological molecules. In this study, the gel pore size was predicted by pulse NMR technique and neural network using a database obtained from gel filtration chromatography and diffusion experiment. Recombinant alkaline phosphatase expressed in insect cells was concentratred 1.5 times by hydrogel ultrafiltration by swelling at 2$0^{\circ}C$ and collapsing at 35$^{\circ}C$ at 53-65% separation efficiency and 78-83% enzyme recovery. Wild and recombinant Autographa californica unclear polyhedrosis viruses (AcNPV) were also concentrated 1.4 and 1.6 times of the feed solution at 48.5 and 60.0% separation efficiency, respectively Hydrogel ultrafiltration appears to be an attractive alternative for the concentration of AcNPV and recombinant proteins from insect cells.

• Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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• 2006.06b
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• pp.451-454
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• 2006

This paper presents an overview of ONP deinking efficiency with cellulolytic enzymes and synthetic collector in alkaline pH. Deinking is a series of unit operations designed to detach ink from cellulose fibers and separate the dispersed ink from the pulp slurry. Deinking chemicals are process aids that enable expensive mill equipment used in these unit operations to operate more efficiently - often much more efficiently. We propose the blended deinking agent with cellulolutic enzymes and synthetic collector in deinking pulp of conventional alkaline method. The deinking efficiency of old news print in alkaline pH was enhanced with enzyme treatments. The brightness of deinked pulp was increased with less residual ink particles and yield of enzymatic deinked pulp was improved compared to the deinked pulp of conventional alkaline method.

• Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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• 2006.11a
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• pp.9-14
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• 2006

Deinking is a series of unit operations designed to detach ink from cellulose fibers and separate the dispersed ink from the pulp slurry. Deinking chemicals are process aids that enable expensive mill equipment used in these unit operations to operate more efficiently. This paper presents an overview of ONP deinking efficiency with cellulolytic enzymes and synthetic collector in low alkalinity process. The deinking efficiency of old news print even in low alkalinity condition was enhanced with enzyme contained complex agent. The brightness of deinked pulp was increased with less residual ink particles and yield of deinked pulp was improved compared to the deinked pulp of conventional alkaline method.

• Korean Journal of Microbiology
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• v.40 no.3
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• pp.254-256
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• 2004

Polymerase chain reaction (PCR) is a powerful tool for precisely amplifying selected DNA sequences that have had a broad impact on genomic studies. When examining human $\alpha$- and $\beta$- tryptase genes which have 95% DNA homology, inconsistent PCR amplification of genomic sequences hampered our progress. This study suggests that long PCR technique on the original DNA digested with restriction enzymes improves both efficiency and sensitivity of PCR. These improved results seem to derived from the effective denaturation of the original genomic DNA template or reduction of formation of secondary structures that block either primer annealing or extension in PCR. Elimination of homo- or hetero-duplex products derived from highly homologous genes provides an additional advantage in this study. This communication describes how the use of restriction enzymes improved these efficiencies, and also facilitated studies of highly homologous genes including tryptase genes.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.84-92
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• 1999

The rumen microbial ecosystem is coming to be recognized as a rich alternative source of genes for industrially useful enzymes. Recent advances in biotechnology are enabling development of novel strategies for effective delivery and enhancement of these gene products. One particularly promising avenue for industrial application of rumen enzymes is as feed supplements for nonruminant and ruminant animal diets. Increasing competition in the livestock industry has forced producers to cut costs by adopting new technologies aimed at increasing production efficiency. Cellulases, xylanases, ${\beta}$-glucanases, pectinases, and phytases have been shown to increase the efficiency of feedstuff utilization (e.g., degradation of cellulose, xylan and ${\beta}$-glucan) and to decrease pollutants (e.g., phytic acid). These enzymes enhance the availability of feed components to the animal and eliminate some of their naturally occurring antinutritional effects. In the past, the cost and inconvenience of enzyme production and delivery has hampered widespread application of this promising technology. Over the last decade, however, advances in recombinant DNA technology have significantly improved microbial production systems. Novel strategies for delivery and enhancement of genes and gene products from the rumen include expression of seed proteins, oleosin proteins in canola and transgenic animals secreting digestive enzymes from the pancreas. Thus, the biotechnological framework is in place to achieve substantial improvements in animal production through enzyme supplementation. On the other hand, the rumen ecosystem provides ongoing enrichment and natural selection of microbes adapted to specific conditions, and represents a virtually untapped resource of novel products such as enzymes, detoxificants and antibiotics.

• The Journal of Internal Korean Medicine
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• v.18 no.1
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• pp.175-190
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• 1997

In order to investigate the effects of Gamihagochosan Extract(加味夏枯草散抽出液) on antitumor effects after human cell lines (A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurement of antitumor effect of Mitomycin C(MMC) and lysosomal enzyme's activities using colony forming efficiency, SRB assay which were regarded as a valuable method for the measurement of antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows : 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 Cells after exposure to the extract of Gamihagochosan extract depressed the growth of tumor cells by concentration of Garnihagochosan. 2. Antitumor activity of the ethanol extract from Gamihggochosan extract and MMC on ascites form of Ehrlich carcinoma in mice is slightly improved. Especially the mean of survival times in the group of 200mg/kg and MMC 0.1mg/kg is improved over 34.9%. 3. When Gamihggochosan extract and MMC are administered together, the weight of tumor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Gamihagochosan extract and MMC are more significantly improved than MMC alone. According to the above result, it could be suggested that Gamihagochosan extract has indirect antitumor effect by the increase of MMC uptake.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1153-1160
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• 2009

Lactosucrose ($4^G-\beta$-D-galactosylsucrose) is an oligosaccharide consisting of galactose, glucose, and fructose. In this study, we prepared lactosucrose from lactose and sucrose using a levansucrase derived from Zymomonas mobilis. Optimum conditions for lactosucrose formation were $23^{\circ}C$, pH 7.0, 18.0% (w/v) lactose monohydrate, and 18% (w/v) sucrose as substrates, and 1 unit of enzyme/ml of reaction mixture. Under these conditions, the lactosucrose conversion efficiency was 28.5%. The product was purified and confirmed to be O-$\beta$-D-galactopyranosyl-($1{\rightarrow}4$)-O-$\beta$)-D-glucopyranosyl-($1{\rightarrow}2$)-$\beta$-D-fructofuranoside, or lactosucrose. A mixed-enzyme system containing a levansucrase and a glucose oxidase was applied in order to increase the efficiency of lactose and sucrose conversion to lactosucrose, which rose to 43.2% as a result.

• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2303-2309
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• 2007

Expressed protein ligation (EPL) technique, joining recombinantly expressed proteins to polypeptides, has been widely adopted for addressing various biological questions and for drug discovery. However, joining two recombinant proteins together is sometimes difficult when proteins are expressed insoluble and unrefoldable, because ligation-active proteins via intein-fusion are obtainable when they are folded correctly. We overcame this limitation coexpressing target protein with additional methionine aminopeptidase (MAP) which enhances removal of the initiation methionine of recombinantly expressed protein. Our approach demonstrated that two domains of 46 kDa 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, a target of herbicide glyphosate, were successfully joined by native chemical ligation, although its C-terminal domain was expressed as an inclusion body. The intein-fused N-terminal fragment of EPSP synthase (EPSPSN, residues 1-237) was expressed and the ligation-active thioester tagged N-terminal fragment (EPSPSN-thioester) was purified using a chitin affinity chromatography and mercapto-ethanesulphonate (MESNA) as intein thiolysis reagent. Its Cterminal fragment (EPSPSC, residues Met237-238CYS-427), expressed as an inclusion body, was prepared from an additional MAP-expressing strain. Protein ligation was initiated by mixing ~1 mM of EPSPSN-thioester with ~2 mM of EPSPSCCYS (residues 238CYS-427). Also we found that addition of 2% thiophenol increased the ligation efficiency via thiol exchange. The ligation efficiency was ~85%. The ligated full-length EPSP synthase was dissolved in 6 M GdHCl and refolded. Circular dichroism (CD) and enzyme activity assay of the purified protein showed that the ligated enzyme has distinct secondary structure and ~115% specific activity compared to those of wild-type EPSP synthase. This work demonstrates rare example of EPL between two recombinantly expressed proteins and also provides hands-on protein engineering protocol for large proteins.