• The Korean Journal of Mycology
• /
• v.20 no.3
• /
• pp.252-258
• /
• 1992

Thirty three species and two species varieties belonging to 14 genera of fungi were collected from 20 cowpea cultivars on glucose Czapek's agar (11 genera and 25 species+1 var.) and glucose-Czapek's agar supplemented with 10% NaCl (7 genera and 18 species+2 var.) at $28{\pm}2^{\circ}C$. The total count of fungi were 6716 colonies/g in all cowpea cultivars. On glucose-Czapek's agar and identified; Aspergillus flavus, A. niger, A. sydowii, A. flavus var. columnaris, A. terreus, Penicillium chrysogenum, Emericella nidutans and Rhizopus stolonifer. The total count of halotolerant or halophilic fungi was 3515 colonies/g on 10% NaCl-glueose-Czapek's agar and identified; the most common species were: A. flavus, A. sydowii, A. tamarii A. flavipes, A. niger, A. flavus var. columnaris, A. ochraceus, A. oryzae and P. chrvsogenusm. Thin layer chrormatographic analysis of chloroform extracts of the different seed samples revealed that four cultivars were naturally contaminated with aflatoxins $B_1,\;B_2,\;G_1$ and $G_2$, $(45-112\;{\mu}g/kg)$.

• KSBB Journal
• /
• v.24 no.1
• /
• pp.30-40
• /
• 2009

Statistical optimization of the production medium was carried out in order to find an optimal medium composition in itaconic acid fermentation process. Itaconic acid utilized in the manufacture of various synthetic resins is a dicarboxylic acid biosynthesized by fungal cells of Aspergillus terreus in a branch of the TCA cycle via decarboxylation of cis-aconitate. Through OFAT (one factor at a time) experiments, six components (glucose, fructose, sucrose, soluble starch, soybean meal and cottonseed flour) were found to have significant effects on itaconic production among various carbon- and nitrogen-sources. Hence, using these six factors, interactive effects were investigated via fractional factorial design, showing that the initial concentrations of sucrose and cottonseed flour should be high for enhanced production of itaconic acid. Furthermore, through full factorial design (FFD) experiments, negative effects of $KH_2PO_4$ and $MgSO_4$ on itaconic acid biosynthesis were demonstrated, when excess amounts of the each component were initially added. Based on the FFD analysis, further statistical experiments were conducted along the steepest ascent path, followed by response surface method (RSM) in order to obtain optimal concentrations of the constituent nutrients. As a result, optimized concentrations of sucrose and cottonseed flour were found to be 90.4g/L and 53.8g/L respectively, with the corresponding production level of itaconic acid to be 4.36 g/L (about 7 fold higher productivity as compared to the previous production medium). From these experimental results, it was assumed that optimum ratio of the constituent carbon (sucrose) and nitrogen (cottonseed flour) sources was one of the most important factors for the enhanced production of itaconic acid.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.33 no.3
• /
• pp.566-570
• /
• 2004

Cultivation conditions for overproduction of lovastatins were investigated from the lovastatin producing strain N-03 which was obtained with NTG (N-methyl-N'-nitro-nitrosoguanidine) treatment from Aspergiliu ferrous ATCC 20542. Produced lactone and acid form of lovastatin were detected, and analyzed by HPLC method. In liquid culture, medium No. 2 containing soy protein produced higher amounts of the lovastatins than medium No. 1 (contained rapeseed oil). In solid culture, maximum production was obtained at 28$^{\circ}C$ for 15 days cultivation using cooked wheat bran. For the overproduction of lovastatin from this strain, solid culture method using plastic bag is more superior than liquid culture.

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.228-232
• /
• 2003

Lovastatin produced by Aspergillus terreus via polyketide pathway is a secondary metabolite with high anti-hypercholesterolemic activity. In this paper we are going to present effective strain development strategies for lovastatin production by comparing the productivity of the mutants obtained through traditional rational screening process and protoplast fusion method. Mutants resistant against various antibiotics and/or antimetabolites showed significantly higher lovastatin productivity than the corresponding mother strains, demonstrating that rational screening method was very efficient in selecting high yielding producers. Recombinant fusants obtained using protoplast fusion between high producers were observed to have very different morphology and physiology as represented by the production and secretion of lovastatin, as well as cell growth pattern. In parallel with the strain development, optimization process for the production medium was carried out in order to find optimal concentrations of the medium components using such a powerful statistical method as response surface method (RSM). It was concluded that not only the optimum production medium but also good morphological characteristics of the high-yielding producers led to higher lovastatin production.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.8
• /
• pp.1084-1091
• /
• 2012

Xylooligosaccharides are functional foods mainly produced during the hydrolysis of xylan by physical, chemical, or enzymatic methods. In this study, production of xylobiose was investigated using oil palm empty fruit bunch fiber (OPEFB) as a source material, by chemical and enzymatic methods. Xylanase-specific xylan hydrolysis followed by xylobiose production was observed. Among different xylanases, xylanase from FXY-1 released maximum xylobiose from pretreated OPEFB fiber, and this fungal strain was identified as Aspergillus terreus and subsequently deposited under the accession Number MTCC- 8661. The imperative role of lignin on xylooligosaccharides enzymatic synthesis was exemplified with the notice of xylobiose production only with delignified material. A maximum 262 mg of xylobiose was produced from 1.0 g of pretreated OPEFB fiber using FXY-1 xylanase (6,200 U/ml) at pH 6.0 and $45^{\circ}C$. At optimized environment, the yield of xylobiose was improved to 78.67 g/100 g (based on xylan in the pretreated OPEFB fiber).

• Microbiology and Biotechnology Letters
• /
• v.22 no.6
• /
• pp.646-650
• /
• 1994

Aspergillus terreus NRRL 1960 was immobilized on various alginate gel beads, Celites, and polyurethane foam cubes, and the comparisons were made for the production of itaconic acid according to the types and sizes of each carrier. The levels of itaconic acid produced from Ca- alginate and Sr-alginate were similar, and the addition of bentonite to Ca- and Sr-alginate resulted in an increase of itaconic acid. The addition of 1.67% bentonite and 0.33% starch to Sr-alginate (SABS bead) produced higher level of itaconic acid (11.59 g/1) than other gel beads. A decrease in the size of Celite increased the itaconic acid production, and the smallest size of Celites, R- 634, produced 6.37 g/l of itaconic acid. Among various types of polyurethane foam cubes, HR 08 (2X2X2 cm) produced about 19 g/l of itaconic acid, which was more efficient than other carriers. In a repeated batch culture using immobilized cells on polyurethane foam cubes (HR 08, 2X2X2 cm), the stability of itaconic acid production was maintained up to 4 batches. Also, the possibility of itaconic acid production by continuous culture was shown in a packed-bed reactor.

• KSBB Journal
• /
• v.22 no.5
• /
• pp.297-304
• /
• 2007

For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

• Microbiology and Biotechnology Letters
• /
• v.44 no.1
• /
• pp.98-105
• /
• 2016

The presence of bacterial strains showing antagonistic activity to common pathogens found in a variety of fermented foods in Korea was explored. A bacterium inhibiting the growth of pathogens such as Aspergillus terreus (KCTC6178), A. flavus (KCTC6984), Staphylococcus aureus (KCCM12214), Escherichia coli O157:H7 (KCCM40406), Bacillus cereus (KCTC1012), Cryptococcus neoformans (ATCC208821), Salmonella typhimurium (ATCC19430), and Listeria monocytogenes (KCTC3569) was isolated from Makgeolli, a Korean traditional rice wine. The strain showing high antipathogenic activity was identified as B. amyloliquefaciens based on the nucleotide sequence of the 16S ribosomal RNA gene. Compared with B. amyloliquefaciens KCTC1660, whose genome has been sequenced, the isolate exhibited significantly low activities of starch-degrading enzymes and high resistance to high temperature and low pH.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.11
• /
• pp.1670-1679
• /
• 2020

The substantial use of fungal enzymes to degrade lignocellulosic plant biomass has widely been attributed to the extensive requirement of powerful enzyme-producing fungal strains. In this study, a two-step screening procedure for finding cellulolytic fungi, involving a miniaturized culture method with shake-flask fermentation, was proposed and demonstrated. We isolated 297 fungal strains from several cellulose-containing samples found in two different locations in Thailand. By using this screening strategy, we then selected 9 fungal strains based on their potential for cellulase production. Through sequence-based identification of these fungal isolates, 4 species in 4 genera were identified: Aspergillus terreus (3 strains: AG466, AG438 and AG499), Penicillium oxalicum (4 strains: AG452, AG496, AG498 and AG559), Talaromyces siamensis (1 strain: AG548) and Trichoderma afroharzianum (1 strain: AG500). After examining their lignocellulose degradation capacity, our data showed that P. oxalicum AG452 exhibited the highest glucose yield after saccharification of pretreated sugarcane trash, cassava pulp and coffee silverskin. In addition, Ta. siamensis AG548 produced the highest glucose yield after hydrolysis of pretreated sugarcane bagasse. Our study demonstrated that the proposed two-step screening strategy can be further applied for discovering potential cellulolytic fungi isolated from various environmental samples. Meanwhile, the fungal strains isolated in this study will prove useful in the bioconversion of agricultural lignocellulosic residues into valuable biotechnological products.

• YAKHAK HOEJI
• /
• v.42 no.5
• /
• pp.473-479
• /
• 1998

Lovastatin (LOVA), a fungal metabolite isolated from cultures of Aspergillus terreus, is a competitive HMG-CoA reductase inhibitor used for the treatment of primary hyper cholesterolemia, and has also been shown to suppress growth in a variety of non-glioma tumor cell lines. A sensitive reversed-phase high-perfonnance liquid chromatographic method with ultraviolet (UV) absorbance detection has been developed to quantitate LOVA in human plasma and urine samples using liquid-liquid extraction procedure. Baseline separation of LOVA and internal standard, simvastatin was achieved on a Novapak $C_{18}$ analytical column with a mobile phase containing 0.025M $NaH_2PO_4$: CAN (35:65, v/v%), adjusted pH to 4.5. The flow rate was set at 1.5ml/min, and the column effluent was monitored by a UV detection at 238nm. The limit of quantification was determined to be 0.5${\mu}$g/ml while extraction efficiency of LOVA ranged from 73.4-82.9% at LOVA concentrations of 0.5 to 10${\mu}$g/ml. Good linearity with correlation coefficients greater than 0.999 was obtained in the range of LOVA concentrations from 0.5 to 10${\mu}$g/ml. The accuracy and the precision were proven excellent with relative standard deviation (RSD, %) and relative error (RE, %) of less than 4.2 and 4.0, respectively. Intraday precision, evaluated at five LOVA concentrations (0.5, 1, 2, 5, 10${\mu}$g/ml) and expressed as RSD ranged from 0-1.82% while the interday precision at the same concentrations ranged from 0.7-10.5%. The analytical method described was then successfully employed for the determination of LOVA concentrations in plasma samples obtained during a phase II clinical trial using high doses of LOVA (30-40mg/kg/day). This method could be further utilized for the ongoing pharmacolkinetic studies and therapeutic drug monitoring of the high-dose LOVA therapy in adenocarcinoma patients.