• Bulletin of the Korean Chemical Society
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• 제31권6호
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• pp.1527-1534
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• 2010

The conventional peptide modification process of guanidination, in which the amino groups of lysine residues are converted to guanidino groups using O-methylisourea to create more basic homoarginine residues, is often used to improve the signal intensity of lysine-containing peptides in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Here, we used three different protease enzymes (trypsin, Lys-C, and Glu-C) to evaluate the effects of guanidination on the MS signals of two enzymatically digested proteins. Horse heart myoglobin and bovine serum albumin were guanidinated either before or after digestion with trypsin, Lys-C, or Glu-C. The resulting peptides were subjected to MALDI-MS, and signal intensities and sequence coverage were systematically evaluated for each digest. Guanidination prior to Glu-C digestion improved sequence coverage for both proteins. For myoglobin, guanidination before enzymatic digestion with trypsin or Lys-C also enhanced sequence coverage, but guanidination after enzymatic digestion enhanced sequence coverage only with Lys-C. For albumin, guanidination either before or after Glu-C digestion increased sequence coverage, whereas pre- or post-digestion guanidination decreased sequence coverage with trypsin and Lys-C. The amino acid composition of a protein appears to be the major factor determining whether guanidination will enhance its MALDI-MS sequence coverage.

• Preventive Nutrition and Food Science
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• 제6권3호
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• pp.180-186
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• 2001

Bone marrow cell proliferating arabinogalactan-like polysaccharide (ALR-3IIa-1-1) has been purified from rhizomes of Atractylodes lancea DC. In order to characterize the essential structure of ALR-3IIa-1-1 for expression of the activity, sequential enzymatic digestion using ego-$\alpha$-L-arabinofurasidase (AFase) and ego-$\beta$-D-(1longrightarrow3)-galactanase (GNase) was employed. After ALR-3IIa-1-1 was digested with the AFase, the GNase digestion cleaved only 10% and 23% of 3-linked and 3,6-branched galactose, respectively, from arabinose-trimmed ALR-3IIa-1-1 (AT-ALR-3IIa-1-1), and gave small amounts of intermediate size (AT-G-2) and shorter oligosaccharides (AT-G-3) fractions in addition to a large amount of the GNase resistant fraction (AT-G-1). When AT-G-1 was redigested gradually with the AFase and GNase, it released trace amounts of oligosaccharides in addition to a large amount of the resistant fraction. When the final enzyme-resistant fraction from AT-G-1 was digested simultaneously with both AFase and GNase, the resistant fraction was significantly degraded into two long fragments (3AT-3G-1 and 2). The mixture of digestion products from the first GNase digestion of AT-ALR-3IIa-1-1 showed a significantly decreased bone marrow cell proliferation activity to about 30% of the activity of ALR-3IIa-1-1, but the GNase resistant fraction (AT-7-1) still had significant activity. Although the second gradual enzymatic digestion of AT-G-1 showed a marginal decrease in activity, the resulting fragments (3AT-3G-1 and 2) by the final simultaneous enzymatic digestion lost most of the activity. Component sugar, methylation and FAB-MS analyses indicated that the digestion products (AT-G-21 AT-G-31 2AT-2G-2 and 2AT-2G-3) released from AT-ALR-3IIa-1-1 by the sequential enzymatic digestion contained galactose-containing oligosaccharides mainly comprising 6-linked galactose, that some of which were partially arabinosylated, and these oligosaccharides were attached to $\beta$-D-(1longrightarrow3)-galactan backbone in its non-reducing terminal side as side chains.

• Food Science and Biotechnology
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• 제16권3호
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• pp.493-495
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• 2007

In this study, the acrylamide contents of foods were estimated via liquid chromatography (LC)/mass spectrometry (MS)/MS after the food matrix constituents had been degraded with digestive enzymes (i.e., pepsin and pancreatin) and extracted with water. The quantities of acrylamide released from samples of cereal, potato chips, peanuts, and coffee were $62{\pm}5.1,\;970,\;106{\pm}20$, and 890 ppb, respectively. No acrylamide was detected in samples of soybean curd (tofu), fish cake, and ham. Compared to the amounts of acrylamide detected after extraction with water only, we noted no significant differences in the soybean curd, fish cake, potato chip, ham, and coffee samples. However, the quantities of acrylamide released from the cereal and peanut samples were approximately 2-fold larger following pretreatment with the digestive enzymes. This study presents a new in vitro enzymatic digestion method which allows for a more accurate estimation of the acrylamide contents of foods.

• Parasites, Hosts and Diseases
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• 제42권2호
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• pp.57-60
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• 2004

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

• Preventive Nutrition and Food Science
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• 제14권1호
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• pp.21-28
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• 2009

In this study, we focused on natural water-soluble antioxidants from Tetraselmis suecica (T. suecica) and Chlorella ellipsoidea (C. ellipsoidea). They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Alcalase, Flavourzyme, Neutrase, and Kojizyme), and the potential antioxidant activity of each was assessed. Most enzymatic digests from T. suecica had a higher radical scavenging activity than those from C. ellipsoidea. Among the enzymatic digests, Kojizyme digest from T. suecica exhibited the highest effect on DPPH radical scavenging. Viscozyme (30.2%) and Neutrase (34.6%) digests from T. suecica exhibited higher hydroxyl radical scavenging activity. Kojizyme digest from T. suecica (81.5%) had strong alkyl radical scavenging activity. Neutrase (61.9%) and Kojizyme (61.5%) digest from T. suecica possessed the highest effects on hydrogen peroxide scavenging. Among the tested samples, Neutrase (TN) and Kojizyme (TK) digests from T. suecica showed the highest antioxidant activity (DPPH, alkyl radical, hydrogen peroxide). Therefore, TN and TK digests were selected for use in the further experiments. Those digests showed enhanced cell viability against $H_2O_2$-induced oxidative damage, and relatively good hydrogen peroxide scavenging activity in an African green monkey kidney (Vero) cell line. These results suggested that an enzymatic digestion will be an effective way for the production of a potential water-soluble antioxidant from a microalgae, T. suecica.

• Asian-Australasian Journal of Animal Sciences
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• 제16권3호
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• pp.417-424
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• 2003

Inhibitory activities against angiotensin I-converting enzyme (ACE) of enzymatic hydrolysates of porcine skeletal muscle proteins were investigated. Myosin B, myosin, actin, tropomyosin, troponin and water-soluble proteins extracted from pork loin were digested by eight kinds of proteases, including pepsin, $\alpha$-chymotrypsin, and trypsin. After digestion, hydrolysates produced from all proteins showed ACE inhibitory activities, and the peptic hydrolysate showed the strongest activity. In the case of myosin B, the molar concentration of peptic hydrolysate required to inhibit 50% of the activity increased gradually as digestion proceeded. The hydrolysates produced by sequential digestion with pepsin and $\alpha$-chymotrypsin, pepsin and trypsin or pepsin and pancreatin showed weaker activities than those by pepsin alone, suggesting that ACE inhibitory peptides from peptic digestion might lose their active sequences after digestion by the second protease. However, the hydrolysates produced by sequential digestion showed stronger activities than those by $\alpha$-chymotrypsin, trypsin or pancreatin alone. These results suggested that the hydrolysates of porcine meat were able to show ACE inhibitory activity, even if they were digested in vivo, and that pork might be a useful source of physiologically functional factors.

• 한국응용곤충학회지
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• 제34권2호
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• pp.95-99
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• 1995

무당벌레(Harmonia axyridis)의 중장 내에서 먹이 단백질이 소화되는 과정을 연구하기 위하여 진딧물과 생간의 AP를 모델 단백질로 이용하였다. 천역 먹이인 긴꼬리볼록진딧물(Megoura crassicauda)과 인공먹이인 닭의 생간은 각각 고유의 acid phosphatase(AP)를 가지고 있으며, 무당벌레의 중장 내부로 들어온 후에도 활성을 나타내었다. 무당벌레의 장에서 자체적으로 생성하여 중장 내부로 분비하는 AP는 관찰되지 않았다. 무당벌레의 단백질 소화력은 먹이의 종류에 따라 차이를 나타내는 경향을 보였다. 생간의 AP는 무당벌레 중장내에서 12시간이 지나면 거의 활성을 잃어 버리나 긴꼬리볼록 진딧물의 AP는 24시간이 경과하여도 강한 활성을 나타내었다.

• Journal of the Korean Wood Science and Technology
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• 제51권4호
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• pp.270-282
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• 2023

Lower termites need symbiotic microbes for cellulose digestion. Elizabethkingia miricola strain BM10 has been proposed as a symbiotic microbe that assists in low-temperature digestion and metabolism of Reticulitermes speratus KMT1, a termite on Bukhan Mountain, Seoul, Korea. In E. miricola strain BM10, β-glucosidase genes expressed at 10℃ were identified, and the psychrophilic enzymatic characteristic was confirmed by heterogeneously expressed proteins. Crude β-glucosidase in the culture broth of E. miricola strain BM10 showed specific enzymatic properties, and its substrate affinity was 4.69 times higher than that of Cellic CTec2. Among the genes proposed as β-glucosidase, two genes, bglB_1 and bglA_2, whose gene expression was more than doubled at 10℃ than at 30℃, were identified. They were heterogeneously expressed in Escherichia coli and identified as psychrophilic enzymes with an optimal reaction temperature of about 20℃-25℃. In this study, E. miricola strain BM10, a symbiotic bacterium of lower termites, produced psychrophilic β-glucosidases that contribute to the spread of the low-temperature habitat of a lower termite, R. speratus KMT1.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.459-462
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• 2000

The pretreatment of used newspaper for the enzymatic digestion preprocess was performed on a percolation reactor and a batch reactor. The test condition of percolation process was $170^{circ}C$, 60min, 1 mL/min, and 400psi, that of batch was $40^{circ}C$, 3hr. and latm Reaction solutions used in pretreatment process were aqueous ammonia, sulfuric acid, water, and hydrogen-peroxide as an oxidizing agent. As a result, the effect of pretreatment was similar to batch and percolation process, but the yield of enzymatic hydrolysis was higher in batch than percolation. This batch pretreatment enhanced enzymatic hydrolysis rate and increased glucose yield from about 15 to 20%. The inhibition factors influenced the rate of enzymatic hydrolysis was investigated, and the ink contented newspaper was the major factor.

• ALGAE
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• 제24권3호
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• pp.169-177
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• 2009

In this study, we focused on natural water-soluble antioxidants from fresh water microalgae, Pediastrum duplex and Dactylococcopsis fascicularis from Jeju Island, Korea. They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase), and the potential antioxidant activity of each was assessed. All enzymatic digests from P. duplex showed significant DPPH scavenging effects. Termamyl (60.6%) digest from P. duplex possessed the highest effects on hydrogen peroxide scavenging. Celluclast (58.1%) and Kojizyme digests (56.9%) from D. fascicularis possessed higher effects on superoxide anion radical scavenging. All enzymatic digests exhibited significant effects on both NO· scavenging and metal chelating. Lipid peroxidation was significantly in inhibited Viscozyme, Termamyl and Kojizyme digests from P. duplex and Ultraflo, Protamex, Kojizyme and Alcalase digests from D. fascicularis. These data suggest that enzymatic digests of the fresh water microalgae, P. duplex and D. fascicularis might be valuable sources of antioxidant which can be applied in food and pharmaceutical industry.