Maximum cellulase production was sought by comparing the activities of the cellulases produced by different Trichoderma reesei strains and Aspergillus niger. Trichoderma reesei Rut-C30 showed higher cellulase activity than other Trichoderma reesei stains and Aspergillus niger that was isolated from soil. By optimizing the cultivation conditions during shake flask culture, higher cellulase production could be achieved. The FP(filter paper) activity of 3.7U/ml and CMCase (Carboxymethylcellulase) activity of 60U/ml were obtained from shake flask culture. When it was grown in 2.5L fermentor, where pH and DO levels are controlled, the enzyme activities were 133.35U/ml (CMCase) and 11.67U/ml(FP), respectively. Ammonium sulfate precipitation method was used to recover enzymes from fermentation broth. The dried cellulase powder showed 3074.9U/g of CMCase activity and 166.7U/g of FP activity with 83.5% CMCase recovery.
Systematic bioconversion process for the production of xylose from agricultural wastes such as barley straw and corn cobs was studied. After the pretreatment in 1 % NaOH solution for 24 hours at 3$0^{\circ}C$, enzymatic hydrolysis of barley straw for 48 hours at 3$0^{\circ}C$ resulted in the liberation of 15.8% of reducing sugar which is equivalent to 87% of total D-xylose content. Among various agricultural wastes, corn cob as well as barley straw was demonstrated to be potent sources for the production of D-xylose by the process of enzymatic conversion.
Acid proteinase has been discovered in Aspergillus niger (acid protease A) and Cryphonectria parasitica (acid proteinase EapC) and it plays major roles in cheese formation from milk. In this study, a partial gene encoding acid proteinase in Penicillium oxalicum HCLF-34 was cloned by using PCR with degenerate primers corresponding to highly conserved regions of the acid proteinase. The partial acid proteinase gene in P. oxalicum HCLF-34 contains an open reading frame of 438 base pairs and encodes an acid proteinase protein of 146 amino aicds. The predicted amino acid sequences showed 71 % homology with acid protease A and 67% homology with EapC.
Background: Otomycosis is a fungal infection that comprises 7~10% of outer ear infections. Although the occurrence is higher in humid climates, relatively few studies have investigated otomycosis occurrences in humid environments. While recurrent chronic otitis media discharge in the ear creates a milieu in which otomycosis is likely to occur, investigations of otomycosis co-occurring with chronic otitis media have been rare. Objective: To examine the characteristics of patients with otomycosis co-occurring with chronic otitis media and identify causative fungi. Methods: The study included 60 patients with chronic otitis media who presented typical otomycosis findings in the outer ear canal and the presence of fungi. Patients were treated in the department of otolaryngology, Daegu Catholic University Medical Center, between July 2011 and June 2018. Results: The mean patient age was 57.77 years, and our study included 20 men and 40 women (p=0.010). The lesion was on the right in 39 patients and on the left in 21 (p=0.020). Ear discharge was the most common chief complaint at diagnosis. Of the 54 patients over age 19, 10 had diabetes (18.5%). Aspergillus was causative in 29 patients and Candida in 31. Aspergillus niger was identified in 15 patients, Aspergillus sp. in 14, Candida parapsilosis in 12, Candida sp. in six, and Candida albicans in five. Conclusion: Otomycosis and chronic otitis media co-occurrences increase with age. The Aspergillus and Candida genera were similar in proportion. A. niger was the most common Aspergillus species, while C. parapsilosis was the most common Candida.
Aspergillus niger B-15 with strong Endo-polygalacturonase (Endo-PG) activities was selected out from a total of 1,573 fungal strains isolated from various testing materials. A mutant strain, U-46, was obtained from the Aspergillus niger B-15 by repeated irradition of ultra-violet light. The objectives of the study were to investigate the fungal properties of the parental and mutant strains obtained and to study the condition of enzyme production and reaction. The results obtained are summarized as follows: 1. The size of conidial head of the U-46 mutant was smaller than that of the parental strains, B-15 and the length of the conidiophore was also shorter than that of the parental strains. 2. The optimum conditions for the Endo-PG production of the parental B-15 strain in the wheat bran Koji were obtained when 40% of water was added to the wheat bran and the temperature was 30 to $35^{\circ}C$. However, the best condition for the mutant U-46 strain was attained when 60 to 70% of water was added and the temperature was $35^{\circ}C$. The optimum growing periods were two to three days for both parental and mutant strains. 3. Under the optimum producing conditions of each strains, the enzymatic activity of the mutant U-46 was 20 times higher than the Endo-PG of the parental strain, B-15. 4. When both strains were cultured in the wheat bran Koji containing 60% of water at $35^{\circ}C$ for three days, the mutant strain. U-46, was about 46 times higher in the Endo-PG activity and about 18 times greater in Exo-PG activity than the parental strain, B-15. The activities of cellulase, $\alpha$-amylase, and glucoamylase were also highly increased in the mutant strain. 5. The mutant strain, U-46, increased its Endo-PG activity up to 20% over that of ordinary case when 1.2 to 1.5% of ammonium sulphate was added to the wheat bran. 6. The optimum condition for Endo-PG activity of crude enzyme of the mutant strain, U-46, was attained when pH of reaction solution was 4.0 to 4.5 and the temperature was $50^{\circ}C$.
This study was conducted to investigate the effects of Aspergillus niger-derived multi-enzyme complex supplementation to feedrestricted lactating sows on performances, milk yield, blood profiles, and manure excretion as compared with ad libitum-fed sows without supplementation of enzyme. Fifty multiparous lactating Berkshire sows were allotted to 5 treatments of 10 sows per treatment during a 28-d lactation period and litter per sow was standardized to 9 suckling piglets. Treatments were ad libitum-fed sows without enzyme and feed-restricted sows supplemented with four increasing levels (0, 0.02, 0.04 and 0.08%) of multi-enzyme complex derived from Aspergillus niger. Blood samples from all sows were collected to determine serum metabolite concentrations before the morning feeding on d 27 of lactation. Litter body weight and a piglet weight at weaning, and litter weight gain significantly (P<0.05) increased with increasing levels of multi-enzyme complex, but there was no significant difference between ad libitum-fed sows without enzyme and feed-restricted sows supplemented with multi-enzyme complex. Body condition score and backfat depth at weaning significantly (P<0.05) increased as multi-enzyme complex level increased. Lactational backfat depth tended (P>0.05) to less decrease with increasing levels of enzyme complex. Serum inorganic phosphorus and non-esterified fatty acid concentrations significantly (P<0.05) increased with increasing levels of enzyme complex. Daily milk yield was not significantly different across treatments, but milk fat yield significantly (P<0.05) increased as multi-enzyme complex level increased. Manure output was significantly (P<0.01) higher for ad libitum-fed sows than for feed-restricted sows, but there was no significant difference among feed-restricted sows supplemented with increasing levels of multi-enzyme complex. Fecal phosphorus amount significantly (P<0.05) decreased with increasing levels of multi-enzyme complex. Feed costs of sows per litter weight gain were reduced by 1.25% to 9.67% with increasing levels of multi-enzyme complex as compared with ad libitum-fed sows without enzyme. The results indicated that multi-enzyme supplementation to feed-restricted lactating sows not only increased litter performances, but also was comparable to ad libitum-fed sows, resulting in reduced feed costs. Moreover, the reduction of fecal phosphorus amount with increasing levels of enzyme complex would contribute to the reduction of environmental pollution.
Kim, Min Sik;Kim, Sinil;Ha, Byeong-Seok;Park, Hye-Young;BaeK, Seong-Yeol;Yeo, Soo-Hwan;Ro, Hyeon-Su
The Korean Journal of Mycology
/
v.42
no.3
/
pp.191-200
/
2014
Nuruk samples collected from various regions in Korea were investigated in terms of fungal contents and diversity. In measurement of colony forming unit (CFU) in Nuruk suspensions on DRBC agar, Nuruk samples MS4, MS8, and MS10 were among the highest fungal density, with $1,278.9{\pm}21.6$ (${\times}10^4$), $1,868.0{\pm}27.7$ (${\times}10^4$), and $775.1{\pm}19.2$ (${\times}10^4$) were among the samples showing the highest fungal density. CFU per 20 mg Nuruk, respectively. The majority of fungal components were yeasts, including Pichia anomala, P. kudriavzevii, Kluyveromyces marxianus, and Saccharomycopsis fibuligera, whereas Aspergillus oryzae and Rhizopus oryzae, the representative Nuruk fungi, were predominant only in the low fungal density Nuruks (MS2, MS5, and MS11). Saccharification capability of the fungal isolates was assessed by measurement of amylase activity in the culture broth. The highest amylase activity was found in A. niger and A. luchuensis, followed by S. fibuligera. A. oryzae and R. oryzae showed fair amylase activity but significantly lower than those of the three fungal species. R. oryzae was suggested to play an additional role in degradation of ${\beta}$-glucan in crop component of Nuruk since R. oryzae was the only fungus that showed ${\beta}$-glucanase activity among the fungal isolates. To confirm the safety of Nuruk, aflatoxigenicity of the isolated Aspergillus was estimated using the DNA markers norB-cypA, aflR, and omtA. All of the isolates turned out to be non-aflatoxigenic as evidenced by the deletion of gene markers, norB-cypA and aflR, and the absence of aflatoxin in the culture supernatants shown by TLC analysis.
Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.
Xiong, Ai Sheng;Yao, Quan-Hong;Peng, Ri-He;Li, Xian;Fan, Hui-Qin;Guo, Mei-Jin;Zhang, Si-Liang
BMB Reports
/
v.37
no.3
/
pp.282-291
/
2004
Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.
Omolaiye, Gabriel Efomeh;Ojo, John Sunday;Oladapo, Michael Ilesanmi;Ayolabi, Elijah A.
Geophysics and Geophysical Exploration
/
v.14
no.1
/
pp.50-57
/
2011
For effective and accurate prediction of overpressure in the Efomeh field, located in the Niger delta basin of Nigeria, integrated seismic and borehole analyses were undertaken. Normal and abnormal pore pressure zones were delineated based on the principle of normal and deviation from normal velocity trends. The transition between the two trends signifies the top of overpressure. The overpressure tops were picked at regular intervals from seismic data using interval velocities obtained by applying Dix's approximation. The accuracy of the predicted overpressure zone was confirmed from the sonic velocity data of the Efomeh 01 well. The variation to the depth of overpressure between the predicted and observed values was less than 10mat the Efomeh 01 well location, with confidence of over 99 per cent. The depth map generated shows that the depth distribution to the top of the overpressure zone of the Efomeh field falls within the sub-sea depth range of 2655${\pm}$2m (2550 ms) to 3720${\pm}$2m (2900 ms). This depth conforms to thick marine shales using the Efomeh 01 composite log. The lower part of the Agbada Formation within the Efomeh field is overpressured and the depth of the top of the overpressure does not follow any time-stratigraphic boundary across the field. Prediction of the top of the overpressure zone within the Efomeh field for potential wells that will total depth beyond 2440m sub-sea is very important for safer drilling practice as well as the prevention of lost circulation.
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