• Microbiology and Biotechnology Letters
• /
• v.41 no.2
• /
• pp.236-241
• /
• 2013

Pilocarpine is an imidazole alkaloid, found exclusively in the Pilocarpus genus, with huge pharmaceutical importance. In order to extract pilocarpine from Pilocarpus jaborandi, environmentally friendly enzyme-assisted extraction was applied. Viscozyme$^{(R)}$ L, a commercially available enzyme cocktail, was used for the study. The conditions for extraction were optimized on the basis of substrates, enzymes, temperatures and pHs. Optimum conditions for extraction with the highest yield were 30 h reaction of 100 mg substance at $45^{\circ}C$ in 40 ml of 50 mM acetic acid, pH 4. A 10% enzyme concentration was found to be the best for extraction. Total pilocarpine content after extraction was analyzed by HPLC. The total pilocarpine content ($1.14{\mu}g/mg$) obtained from Viscozyme$^{(R)}$ L treatment was 3.08-fold greater than those of the control treatment ($0.37{\mu}g/mg$).

• KSBB Journal
• /
• v.28 no.2
• /
• pp.65-73
• /
• 2013

This study aimed to investigate the enzymatic hydrolysis of mannitol using Viscozyme$^{(R)}$ L, Celluclast$^{(R)}$ 1.5 L, Saczyme$^{(R)}$, Novozym$^{(R)}$, Fungamyl$^{(R)}$ 800 L, Driselase$^{(R)}$ Basidiomycetes sp., and Alginate Lyase, and to optimize of reaction conditions for production of reducing sugar. Response surface methodology (RSM) based on central composite rotatable design was used to study effects of the independent variables such as enzyme (1-9% v/w), reaction time (10-30 h), pH (3.0-7.0) and reaction temperature ($30-70^{\circ}C$) on production of reducing sugar from mannitol. The coefficient of determination ($R^2$) of $Y_1$ (yield of reducing sugar by Viscozyme$^{(R)}$ L) and $Y_3$ (yield of reducing sugar by Saczyme$^{(R)}$) for the dependent variable regression equation was analyzed as 0.985 and 0.814. And the p-value of $Y_1$ and $Y_3$ showing 0.000 and 0.001 within 1% (p < 0.01), respectively, was very significant. The optimum conditions for production of reducing sugar with Viscozyme$^{(R)}$ L were 9.0 % (v/w) amount of enzyme, 30.0 hours of reaction time, pH 4.5 and $30.0^{\circ}C$ of reaction temperature, and those with Saczyme$^{(R)}$ were 9.0% (v/w) of amount of enzyme dosage, 30.0 h of reaction time, pH 7.0 and $30.0^{\circ}C$ of reaction temperature, consequently, the predicted reducing sugar yields were 22.5 and 27.9 mg/g-mannitol, respectively.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.7
• /
• pp.1233-1241
• /
• 2017

The ginsenoside Rh2 has strong anti-cancer, anti-inflammatory, and anti-diabetic effects. However, the application of ginsenoside Rh2 is restricted because of the small amounts found in Korean white and red ginsengs. To enhance the production of ginsenoside Rh2-MIX (comprising 20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3 as a 10-g unit) with high specificity, yield, and purity, a new combination of enzymatic conversion using the commercial enzyme Viscozyme L followed by acid treatment was developed. Viscozyme L treatment at pH 5.0 and $50^{\circ}C$ was used initially to transform the major ginsenosides Rb1, Rb2, Rc, and Rd into ginsenoside F2, followed by acid-heat treatment using citric acid 2% (w/v) at pH 2.0 and $121^{\circ}C$ for 15 min. Scale-up production in a 10-L jar fermenter, using 60 g of the protopanaxadiol-type ginsenoside mixture from ginseng roots, produced 24 g of ginsenoside Rh2-MIX. Using 2 g of Rh2-MIX, 131 mg of 20(S)-Rh2, 58 mg of 20(R)-Rh2, 47 mg of Rk2, and 26 mg of Rh3 were obtained at over 98% chromatographic purity. Then, the anti-cancer effect of the four purified ginsenosides was investigated on B16F10, MDA-MB-231, and HuH-7 cell lines. As a result, these four rare ginsenosides markedly inhibited the growth of the cancer cell lines. These results suggested that rare ginsenoside Rh2-MIX could be exploited to prepare an anti-cancer supplement in the functional food and pharmaceutical industries.

• KSBB Journal
• /
• v.30 no.2
• /
• pp.53-57
• /
• 2015

In this study, the production of total reducing sugar from macro green-algae Enteromorpha intestinalis by enzymatic hydrolysis was investigated. As a result of enzymatic hydrolysis using 13 kind commercial enzymes, the highest yield of 8.75% was obtained from Viscozyme L, which is multi-enzyme complex such as cellulase, arabanase, beta-glucanase, hemicellulase and xylanase. As a control, only 0.33% and 0.27% yield were obtained from 1% sulfuric acid and 0.05 M citrate buffer (pH 4.8), respectively. In the case of enzyme mixture, the mixture of $Viscozyme^{(R)}$ L and $Cellic^{(R)}$ CTec2 (1:1) was presented the highest yield of 10.67%. Finally, the 14.99% yield was obtained at 36 hr under the condition of 10% biomass and 30% enzyme mixture.

• Korean journal of food and cookery science
• /
• v.26 no.1
• /
• pp.18-25
• /
• 2010

This study was carried out to investigate the total polyphenol and flavonoid contents and antioxidative activities of enzymatic digests from dried Citrus unshiu and C. grandis peels. The yields of digests from dried C. unshiu and C. grandis peels were high in viscozyme (a carbohydrase) and kojizyme (a protease), and enzymatic digests from dried C. grandis peels appeared highly comparable to those of C. unshiu. Total polyphenol contents were high in ultaflo (a carbohydrase) and alcalase and flavourzyme (proteases), and the digests from dried C. unshiu peels appeared high in comparison to C. grandis. Total flavonoid contents were high in ultaflo, alcalase, and water extract. DPPH radical scavenging activities appeared very high in digests from dried C. grandis peels in comparison to C. unshiu, and was the highest in viscozyme and kojizyme. The viscozyme digest displayed particularly high activity. Hydrogen peroxide scavenging activities increased somewhat with increasing amounts of digests, but displayed very high activity, more than 91%, except kojizyme the digest from dried C. unshiu peel, at 2.0 mg/mL. Alkyl radical scavenging activities increased rapidly with increasing amounts of digest, and all enzyme digests from dried C. unshiu and C. grandis peels displayed very high activities at more than 0.5 mg/mL. Hydroxyl radical scavenging activities increased rapidly with increasing amounts of digests, and all enzymatic digests from dried C. unshiu and C. grandis peels displayed relatively low activities in comparison to other activated oxygen species.

• Journal of Applied Biological Chemistry
• /
• v.61 no.1
• /
• pp.101-108
• /
• 2018

This study was conducted to establish optimized ${\beta}-glucan$ extraction method through enzymatic hydrolysis from Phellinus baumii and investigate ${\beta}-glucan$ contents and physicochemical properties. The optimal condition was obtained with the enzyme concentration of 0.66% (v/v), reaction time of 6.08 h ($R^2=0.9245$) and the ${\beta}-glucan$ contents from the Phellinus baumii extracts under the optimized condition was 1.9594 g/100 g. ${\beta}-Glucan$ yield (0.76-16.40%) of enzyme beta-glucan extract (EBE) was three fold higher than that of non-enzyme beta-glucan extract (NEBE). ${\beta}-Glucan$ purity (11.15-59.05%) of non-enzyme beta-glucan (NEB) and that of enzyme beta-glucan (EB) were higher than that of NEBE and that of EBE. ${\beta}-Glucan$ purity of EB (59.05%) and ${\beta}-glucan$ contents of EB (3.38 g/100 g) showed higher than those of others. Total sugar contents (0.61-1.17 mg/mL) showed that NEB and EB were higher than that of NEBE and EBE, EB had the highest total sugar content as 1.17 mg/mL, respectively. Protein contents (0.44-11.73 mg/mL) of NEBE and that of EBE were higher than that of NEB, that of EB. In FT-IR spectrum, the band at $890cm^{-1}$ of microcapsule was attributed to a ${\beta}-1,3-glucan$. The toxicities of ${\beta}-glucan$ from Phellinus baumii in both melanoma cell lines was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoli um bromide assay and ${\beta}-glucan$ from Phellinus baumii has no toxicity until $30{\mu}g/mL$. The effects of ${\beta}-glucan$ from Phellinus baumii on inhibition of cancer cell proliferation were detected by using a wound healing assay. The effect of NEB and EB were higher than NEBE and EBE, especially $30{\mu}g/mL$ of EB had the highest in both melanoma cell lines.

• KSBB Journal
• /
• v.25 no.3
• /
• pp.271-276
• /
• 2010

This study was carried out to obtain the basic information for isoflavonoid contents that can be used to index Astragalus membranaceus B. cultivated in the Republic of Korea and China. Isoflavonoid contents in Astragalus membranaceus B. which were cultivated in various areas (Jecheon, Jeongseon, Yeongju, and Taebaek in Korea, and China) and ages (1-year-old, 3-years-old) were determined. Calycosin and formononetin as major constituents were determinated by HPLC method in Astragalus membranaceus B. The results show that there were no statistically significant differences for the average contents of isoflavonoids among 1-year-old and 3-years-old. However, isoflavonoid contents were significant differences according to the cultivation areas. HPLC analysis showed that the calycosin content of 1-year-old at Jeongseon was the highest level of $0.090{\pm}0.002%$ and that of 1-year-old at Yeongju was the lowest level of $0.010{\pm}0.001%$. The highest level of formononetin content was $0.050{\pm}0.001%$ of 1-year-old at China, while the lowest level was $0.020{\pm}0.001%$ of 1-year-old at Yeongju. These results strongly suggest that contents of isoflavonoid in Astragalus membranaceus B. might be quite different with respect to the cultivation areas.