This study was activity of ${\alpha}-amylase$, glucoamylase of liquid Nuruk prepared using liquid Nuruk (NK) and Aspergillus kawachii (AK), Aspergillus niger (AN), Aspergillus oryzae (AO), Monascus kaoliang (MK). To investigate the relationship between the enzymatic activity and the total sugar content of liquid Nuruk depending on the types of Nuruk molds and the degree of rice-polishing. The activity of ${\alpha}-amylase$ depending on the types of Nuruk molds was shown to be 8.82, 8.72 units/mL in AN and AK treatments in brown rice liquid Nuruk at 24 hours after incubation, as the degree of rice-polishing increased, the activity of ${\alpha}-amylase$ was significantly lower (p<0.05). When brown rice was incubated in AN, it showed 8.83 units/mL at 48 hours after incubation, which was the highest activity, but there was no significantly difference (p<0.05), as the degree of rice- polishing was higher, the activity of ${\alpha}-amylase$ was lower. The activity of glucoamylase depending on the degree of rice-polishing showed 3,013 units/mL in AO treatment in brown rice liquid Nuruk at 24 hours after incubation, and the enzymatic activity was significantly higher (p<0.05). As the degree of rice-polishing increased, the activity of glucoamylase decreased, so liquid brown rice Nuruk showed the highest enzymatic activity, liquid white rice Nuruk was the lowest enzymatic activity. The highest enzymatic activity appeared in liquid Nuruk with brown rice at 48 hours after incubation. The activity of ${\alpha}-amylase$, glucoamylase showed higher enzymatic productivity as the degree of rice-polishing was lower, and there was an inverse correlation with the total sugar content.
Industrial strain improvement is concerned with developing or modifying microorganisms used in production of commercially important fermentation products. The aim is to reduce the production cost by improving productivity of a strain and manipulating specific characteristics such as the ability to utilize cheaper raw materials or resist bacteriophages. The traditional empirical approach to strain improvement is mutation combined with selection and breeding techniques. It is still used by us to improve the productivity of organisms in amino acids, organic acids and enzymes production. The breeding of high L-lysine-producing strain Au112 is one of the outstanding examples of this approach. It is a homoserine auxotroph with AEC, TA double metabolic analogue resistant markers. The yield reaches 100 g/l. Besides, the citric acid-producing organism Aspergillus niger, Co827, its productivity reaches the advanced level in the world, is also the result of a series mutations especially with $^60Co{\gamma}$-radiation. The thermostable .alpha.-amylase producing strain A 4041 is the third example. By combining physical and chemical mutations, the strain A 4041 becomes an asporogenous, catabolite derepressed mutant with rifamycin resistant and methionine, arginine auxotroph markers. The .alpha.-amylase activity reaches 200 units/ml. The fourth successful example of mutation in strain improvement is the glucoamylase-producing strain Aspergillus niger SP56, its enzyme activity is 20,000 units/ml, 4 times of that of the parental strain UV-11. Recently, recombinant DNA approach provides a worthwhile alternative strategy to industrial strain improvement. This technique had been used by us to increase the thermostable .alpha.-amylase production and on some genetic researches.
Industrial strain Improvement is concerned with developing or modifying microorga-nisms used In production of commercially important fermentation products. The aim is to reduce the production cost by improving productivity of a strain and manipulating specific cilarafteristic such as the ability to utilize cheaper raw materials or resist bacteriophages. The traditional empiri-cal approach to strain improvement is mutation combined with selection and breeding techniques. It is still used by us to improve the productivity of organisms in amino acids. organic acids andenzymes production. The breeding of high L-lysine-producing strain Au112 is one of the outstanding examples of this approach. It is it homoserine auxotroph with AEC, TA double metabolicanalogue resistant markers. The yield reaches 100g/1. Resides, the citric acid-producing organism Aspergillus nuger, Co827, its productivity reches the advanced level in the world, is also the result of a series mutations expecially with Co Y-radiation. The thermostable a-amylaseroducing strain A 4041 is the third example. By combining physical and chemical multations. the strain ,A 4041becomes an asporogenous, catabolite derepressed mutant with rifamycin resistant and methionine, arginine auxotroph markers. The a-amylase activity reaches 200 units/ml. The fourth successful example of mutation in strain improvement is the glucoamylase-producing strain Aspergillus nigerSP56 its enzyme activity is 20,000 units/ml, 4 times of that of the parental strain UV_11. Recently recombinant DNA approach Provides a worth while alternative strategy to Industrial strain improve-ment. This technique had been used by us to increase the thermostable a-amylase production and on some genetic researches.
The bioethanol for use as a liquid fuel by fermentation of renewable biomass as an alternative to petroleum is important from the viewpoint of global environmental protection. Recently, many scientists have attempted to increase the productivity of bioethanol process by developing specific microorganism as well as optimizing the process conditions. In the present study, which is based on our previous investigation on the pretreatment process, theproductivity of bioethanol obtained from simultaneous saccharification and fermentation (SSF) process was compared between various domestic materials including barley, brown rice, corn and sweet potato. Additionally, Solid glucoamylase (SGA; developed in Changhae Co.), from modified strain with UV, was used. The result was compared to commercial glucoamylase (GA). It was observed that the fermentation rate was increased together with the yield which can be derived from the final ethanol concentration. Especially, in the case of brown rice, compared to the experimental results using GA, the final ethanol concentration was 1.25 times higher and 18.4 g/L of the yield was increased. Also, the time required for reaching 95% of the maximum ethanol concentration is significantly reduced, which is approximately 36 hours, compared to 88 hours using GA. It means that SGA has excellent saccharogenic power.
Journal of the Korean Society of Food Science and Nutrition
/
v.29
no.6
/
pp.987-994
/
2000
Aspergillus coreanus NR 15-1, Asp. oryzae NR 15-3 and Asp. oryzae NR 2-5 isolated from traditional Korean nuruk were screened as parental strains producing starch hydrolyzing enzymes. They were mutagenized by N-methyl -N'-nitro-N-nitrosoguanidine (NTG) and mutants were isolated for analysis of various amylase activities and the ability of acid production. Among them, the mutants harboring high saccharogenic activity, dextrinogenic activity, and the ability of acid production were selected. Fifteen, six, and five strains of mutants were isolated from Asp. coreanus NR 15-1, Asp. oryzae NR 2-5, and Asp. oryzae NR 15-3, respectively followed by NTG mutagenesis. Among these mutants, thirteen strains were identified as auxotrophic mutants. \ulcorner (Arg. ̄) mutant from Asp. coreanus NR 15-1 showed high glucoamylase activity and total acid productivity. Z6 (Ade. ̄) mutant from Asp. oryzae NR 2-5 showed the highest $\alpha$-amylase activity, therefore \ulcorner and Z6 mutant were selected.
We investigated the possibility of industrial application and economit process of high temperature fermentation by thermotolerant alcohol producing yeasts as previously reported. From the 20% glucose media, the RA-74-2 produced 11.8% (v/v) ethanol at $32^{\circ}C$ (0.5% inoculum) and 10.6% (v/v) ethanol at $40^{\circ}C$ (3% inoculum), respectively. Also, 11.3% (v/v) ethanol was produced for 96 hours in the temperature-gradient fermentation. These results suggest that the RA-74-2 could isuccessfully be applied to save the cooling water and energy in industrial scale without re-investment or modification of established fermentation systems. When potato starch was used as the substrate for the RA-74-2, high temperature fermentation above $40^{\circ}C$ was more appropriate for industrial utilization because organic nitrogen was not necessary to economical fermentation. As the naked barley media just prior to industrial inoculation, taken from the Poongkuk alcohol industry Co., were used, 9.6% (v/v) ethanol was produced at $40^{\circ}C$ for 48 hours in jar-fermentor scale (actually, 9.5-9.8% (v/v) ethanol was produced at 30~$32^{\circ}C$ for 100 hours in industrial scale). The ethanol productivity was increased by the high glucoamylase activity as well as the high metabolic ratio at $40^{\circ}C$ Therefore, if the thermotolerant yeast RA-74-2 would be used in industrial scale, we could obtain a high productivity and saving of the cooling water and energy. Meanwhile, the RA-912 produced 6%(v/v) ethanol in 10% glucose media at $45^{\circ}C$ and showed the less ethanol-tolerance compared with industrial strains. As the produced alcohol was recovered by the vacuum evaporator at $45^{\circ}C$ in 15% glucose media, the final fermentation ratio was enhanced (76% of theoretical yields). This suggest that a hyperproductive process could be achieved by a continuous input of the substrate and continuous recovery of the product under vacuum in high cell-density culture.
Shin, Gyeong Yeon;Choi, Hye Jung;Kang, Yang-Rae;Nam, Ki-Du;Song, Ju Yeong;Joo, Woo Hong
Microbiology and Biotechnology Letters
/
v.45
no.2
/
pp.162-167
/
2017
For the production of bioethanol by the synchronous saccharification and fermentation (SSF) process, bio-capsule formation was attempted. Many saccharifying fungal strains and fermentative yeast strains were first screened. Aspergillus sp. BCNU 6200, Penicillium sp. BCNU 6201, and P. chrysogenum KACC 44363 were found to be excellent producers of saccharifying enzymes such as ${\alpha}$-amylase and glucoamylase. Saccharomyces cerevisiae IFO-M-07 showed the highest ethanol productivity among the tested strains. Secondly, we determined the optimal conditions for pellet formation, and those for bio-capsule formation. All the tested fungal strains formed pellets, and the optimal conditions for bio-capsule formation were $28^{\circ}C$ and 120 rpm. Lastly, SSF process was performed using a bio-capsule. An ethanol yield of 3.9% was achieved by using the Aspergillus sp. BCNU 6200 bio-capsule (Aspergillus sp. BCNU 6200 + S. cerevisiae IFO-M-07) at $30^{\circ}C$ with shaking at 120 rpm during the 10 days of incubation. The results provide useful information on the application of a bio-capsule in bioethanol production under the SSF process.
Kim, Hyun-Soo;Hyun, Ji-Sook;Kim, Jung;Ha, Hyun-Pal;Yoo, Dae-Sik
Microbiology and Biotechnology Letters
/
v.26
no.5
/
pp.456-464
/
1998
For the standardization and quality improvement of traditional Korean Nuruk, 10 strains of fungi, which were isolated from Nuruks and showed good productivity of the saccharogenic and dextrinogenic enzymes, acid and flavor, were selected and their enzymological characteristics and identification were carried out. Aspergillus spp. and Rhizopus sp. showed a high liquefying activity without regard to cultivation time, whereas the majority of strains except for Rhizopus sp. had decreasing saccharifying activity in proportion to the increase in cultivation time. Aspergillus spp. No.17-2, No.17-6 and Rhizopus sp. No.18-1 showed high liquefying and saccharifying activity after 15 and 30 day cultivation. The optimum temperature of most of these saccharogenic and dextrinogenic enzymes was from 40$^{\circ}C$ to 60$^{\circ}C$, and their optimum pH was extensive between pH 3 and pH 11. But Penicillium spp.(2 strains) and Rhizopus sp. showed low activity under the alkalic and acidic conditions. Among these isolated strains, 5 strains which had shown the high productivity of materials were identified as Aspergillus oryzae NR3-6 and Aspergillus oryzae NR17-6, Aspergillus penicilloides NR12-1, Penicillium expansum NR7-7 and Rhizopus oryzee NRl8-1, respectively. Five kinds of mixed culture were carried out and all of them showed a better productivity of saccharogenic and dextrinogenic enzymes than single culture. These results indicate that it is possible to make traditional Korean liquors of good quality by using these fungi.
Microorganisms capable of degrading the raw naked barley were isolated from soil, and the amylase productivity of each strain was examined on plate contained 2% raw naked barley. Of the fungi and actinomycetes tested, 71 strains were subjected to subsequent testing for amylase production, and 4 strains were selected as potent amylase producers. Among them, Strain No. 281 produced the most potent raw naked barley saccharifying enzyme, and was identified as genus Rhizopus from morphological and physiological studies. The ratio of raw starch saccharifying activity (RDA) of the crude enzyme derived from the Rhizopus sp. No. 281 was showed 2-3 fold higher than that of commercial enzyme when the raw naked barley was used as the substrate. In the case of uncooked alcohol fermentation using Rhizopus sp. No. 281 glucoamylase preparation, the alcohol yield of the broth was 2% higher than that of the commercial enzyme.
Mutational experiments were performed to imporve the cellulase productivity of Aspergillus phoenicis KU175, isolated from the southern part of Korea, as a high cellulase producer. By treatment ultra-violet light nad 4-NQO(4-Nitroquinoline-N-Oxide), mutation waas induced, and treatment ultra-violet light and 4-NQO (4-Nitroquinoline-N-Oxide), mutation was induced, and A.phoenicis KU175-115 was finally selected for its highest avicelase production. Avicelase production of the mutant was increased about 2 times compared with those of the wild strain. However, activities of other hydrolytic enzymes, such as amylase, protease and nuclease, of the mutant strain didn't show a marked difference compared with those of the nuclease, of the mutant strain didn't show a marked difference compared with the wild strain, except slight increase in ribonuclease activity and slight decrease in glucoamylase activity. Avicelases from the mutant strain selected were purified from wheat bran culture by successive salting out, followed by dialysis and column chromatography, and their charcteristics were compared with thosw of the wild strain. Avicelase was separated into three peaks in the mutant strain as well as in the case of wild strain. Avicelase II activity of the mutant strain was prominently higher than that of the wild strain, while avicelase I and III activities of those were equivalent. The optimal pH ranges and stability of avicelase II from the mutant strain were pH4-5 and pH3.5-6.0, respectively, as well as in the case of the wild strain. The optimal temperature and thermal stability of avicelase II from the mutant strain were $40{\sim}50^{\circ}C\;and\;20{\sim}55^{\circ}C$, respectively. These results were same as those of the wild strain. By the using of Eadie-Hofastee plot, $K_m\;and\;V_{max}$ of avicelase II from the mutant and the wild strain were calculated to be 2.29mg/ml and $4.84{\mu}g$ reducing sugar as glucose per min equally, from the line fitted to the data by the least square method. Activity of avicelase II from the mutant strain was slightly activated by $Mg^{++}\;but\;inhibited\;by\;Cu^{++}, \;Mn^{++}\;and\;Zn^{++}$, as well as in the case of the wild strain. Therefore, it was concluded that the mutant didn't induce the formation of another avicelase isozyme, or the changes in the properties of avicelase, but induce the changes in the productively of the same avicelase II by the action of regulatory gane.
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