Mesut Taskin

Asymmetric reduction of substituted acetophenones using once immobilized Rhodotorula glutinis cells

The asymmetric reductions of acetophenone and its analogues using once immobilized Rhodotorula glutinis cells were studied. The performance and reaction parameters of the immobilized cells were also investigated and it was determined that the cells could be used 15 times in batch processes. All chiral alcohols obtained using purification procedures were of sufficient enantiopurity (>99%) of the (S)-enantiomer. The applicability of the optimized process for a preparative scale bioreduction was shown. Under the optimum conditions, 35mM (4.3g) of the product ((S)-1-phenylethanol) was produced from 45mM (5.4g) of the substrate (acetophenone) with one time immobilized R. glutinis EBK-2 cells (6g wet weight). The yield was calculated as 77%. In this study, it was found that the buffer level had a very significant effect on the reaction activity. Our results demonstrate that the optimized process can be implemented on a preparative scale.

Production of carotenoids by Rhodotorula glutinis MT-5 in submerged fermentation using the extract from waste loquat kernels as substrate

BACKGROUND The aim of present study was to investigate the feasibility of the hydrolysate extracts from waste loquat kernels as substrate in submerged culture of yeast Rhodotorula glutinis MT-5 for carotenoid production. RESULTS Loquat kernel was found to have high protein (22.5%) and total carbohydrate (71.2%) contents. Dried and powdered loquat kernels were subjected to acid hydrolysis with 2 mol L⁻¹ HCl. The hydrolysate obtained was used for the preparation of loquat kernel extract and detoxified loquat kernel extract. The detoxification of hydrolysate was performed with Ca(OH)₂. Among the 10 R. glutinis isolates, the MT-5 was found to be best in order to produce carotenoid using the extract as substrate. Production media prepared with detoxified loquat kernel extract or loquat kernel extract gave maximum biomass concentrations of 12.64 and 11.37 g L⁻¹, and maximum carotenoid concentrations of 72.36 and 62.73 mg L⁻¹, respectively. CONCLUSION This study has provided effective processes for the conversion of waste material of plant origin to the extracts which are very rich in term of total fermentable sugar. The practicability of the extracts as fermentation substrate was proven in carotenoid production. To the best of our knowledge, this is the first report on use of this waste material as a substrate in yeast fermentations.

Reactive dye bioaccumulation by fungus Aspergillus niger isolated from the effluent of sugar fabric-contaminated soil

The present study dealt with the decolorization of textile dye Reactive Black-5 by actively growing mycelium of Aspergillus niger MT-1 in molasses medium. It was found that the fungus, which was isolated from the effluent of sugar fabric-contaminated soil, was capable of decolorizing the Reactive Black-5 dye in a wide range of temperature, shaking speed and pH values. The experiments also revealed that highest dye decolorization efficiency was achieved with cheap carbon (molasses sucrose) and nitrogen (ammonium chloride) sources. Under the optimized culture conditions, the complete decolorization (100%) of 0.1 g/L dye was achieved in 60 hours. The dominant mechanism of dye removal by the fungus was found to be probably bioaccumulation. Fungal growth in small uniform pellet form was found to be better for dye bioacumulation. Molass as carbon source increased dye bioaccumulation by stimulating the mycelial growth in small uniform pellet form. The maximum bioaccumulation efficiency of fungus for dye was 91% (0.273 g bioaccumulated dye) at an initial dye concentration of 0.3 g/L in 100 hours. It was shown for the first time in the present study that the effluent of sugar fabric-contaminated soil was a good source of microorganisms, being capable of decolorizing snythetic textile dyes.

Effects of extremely low magnetic field on the production of invertase by Rhodotorula glutinis

Invertase is an important enzyme used in many fields especially in food industry to produce fructose syrups. The current study focused on increasing invertase production by exposing Rhodotorula glutinis to extremely low magnetic fields (ELMF; 0 and 7 mT). For this purpose, the microorganism was allowed to grow in normal magnetic field and ELMF for 72 hours at the same temperature (24 ± 2°C). The fermentation was carried out in submerged culture for 120 hours. The results showed that invertase production is strongly dependent on the growth conditions of the microorganism. Both of the different magnetic fields applied to R. glutinis increased invertase production ranged from 48%-67% when compared with the control. On the other hand, ELMF treatment increased biomass formation about 14%-28% when compared with the control. As a result, magnetic field treatment could effectively be used in the production of invertase by R. glutinis.

A new strategy for improved glutathione production from Saccharomyces cerevisiae: use of cysteine‐ and glycine‐rich chicken feather protein hydrolysate as a new cheap substrate

BACKGROUND Glutathione (GSH) is composed of the amino acids glutamic acid, cysteine and glycine. This study investigated the usability of chicken feather protein hydrolysate (chicken feather peptone, CFP) as a substrate for GSH production from Saccharomyces cerevisiae. RESULTS CFP was found to be rich in ash (36.7 g per 100 g), protein (61.1 g per 100 g) and minerals (S, P, K, Ca, Fe, Na and Mg). It also had high contents of cysteine and glycine. CFP augmented biomass and GSH production by 53 and 115% respectively compared with the control medium. The highest biomass (17.4 g l(-1)) and GSH (271 mg L(-1)) concentrations were attained in CFP medium. The second highest biomass (16.8 g l(-1)) and GSH (255 mg L(-1)) concentrations were obtained in fish peptone medium. It was assumed that the high mineral, cysteine and glycine contents of CFP were related to cell growth and GSH synthesis in S. cerevisiae. CONCLUSION This is the first report on the effect of cysteine- and glycine-rich protein hydrolysates on GSH production from S. cerevisiae. In this regard, CFP was tested for the first time as a GSH production substrate. As an additional contribution, a new hydrolysis process was developed for the preparation of protein hydrolysates.

Utilization of chicken feather hydrolysate as a novel fermentation substrate for production of exopolysaccharide and mycelial biomass from edible mushroom Morchella esculenta.

This study was performed to investigate the usability of chicken feather hydrolysate (Chicken feather peptone (CFP)) as substrate for mycelial biomass and extracellular polysaccharides (EPS) production from edible mushroom Morchella esculenta. The ability of CFP to support biomass and EPS production in edible mushroom M. esculenta was compared to those of two commercial peptones (Tryptone peptone (TP) and Fish peptone (FP)). The maximum biomass (16.3 g/l) and EPS (4.8 g/l) concentrations were achieved with TP. Second, high biomass (15.9 g/l) and EPS (4.6 g/l) concentrations were obtained with CFP. Also, biomass and EPS concentrations in CFP medium were statistically near to those in the TP medium. CFP and TP resulted in not only uniform pellets with smaller size (5 mm) but also faster mycelial growth compared to FP. This study showed for the first time that CFP could be effectively used as a novel EPS production substrate.

Production of (R)-1-phenylethanols through bioreduction of acetophenones by a new fungus isolate Trichothecium roseum.

A total of 120 fungal strains were isolated from soil samples and evaluated in the bioreduction of substituted acetophenones to the corresponding (R)-alcohols. Among these strains, isolate Trichothecium roseum EBK-18 was highly effective in the production of (R)-alcohols with excellent enantioselectivity (ee > 99%). Gram scale preparation of (R)-1-phenylethanol is reported.

ENHANCEMENT OF INVERTASE PRODUCTION BY Aspergillus niger OZ-3 USING LOW-INTENSITY STATIC MAGNETIC FIELDS

The aim of this study is to investigate the effect of low-intensity static magnetic fields (SMFs) on invertase activity and growth on different newly identified molds. The most positive effect of SMFs on invertase activity and growth was observed for Aspergillus niger OZ-3. The submerged production of invertase was performed with the spores obtained at the different exposure times (120, 144, 168, and 196 hr) and magnetic field intensities (0.45, 3, 5, 7, and 9 mT). The normal magnetic field of the laboratory was assayed as 0.45 mT (control). Optimization of magnetic field intensity and exposure time significantly increased biomass production and invertase activity compared to 0.45 mT. The maximum invertase activity (51.14 U/mL) and biomass concentration (4.36 g/L) were achieved with the spores obtained at the 144 hr exposure time and 5 mT magnetic field intensity. The effect of low-intensity static magnetic fields (SMFs) on invertase activities of molds was investigated for the first time in the present study. As an additional contribution, a new hyper-invertase-producing mold strain was isolated. Supplemental materials are available for this article. Go to the publishers online edition of Preparative Biochemistry and Biotechnology to view the supplemental file.

Efficient Synthesis of （S）-1-（2-chlorophenyl）ethanol in the Submerged Culture of Alternaria alternata Isolate

Abstract (S)-1-(2-chlorophenyl)ethanol is a key intermediate of L-cloprenaline used for relieving asthma symptoms. The asymmetric reduction of 2-chloroacetophenone 1 to (S)-1-(2-chlorophenyl)ethanol 2 in the submerged culture of Alternaria alternata isolates was studied. A. alternata EBK-8 isolate was the most effective biocatalyst. The bioactivity of the fungus could be significantly improved by the optimization of culture conditions. Parameters such as pH, temperature, agitation, and incubation time considerably influenced the substrate conversion and the optical purity of the product. The reaction was carried out in a culture medium at a substrate concentration of 30 mmol/L and produced the desired product with high conversion (100%) and isolated yield (80%) with an excellent enantiomeric excess (ee) of >99%. Under the optimum conditions, after 54 h reaction time, 24 mmol/L 2 from 30 mmol/L 1 could be produced. As a result, the submerged fermentation of A. alternata EBK-8 was found to be suitable for the asymmetric reduction of 1 to 2.

Inulinase production by Geotrichum candidum OC-7 using migratory locusts as a new substrate and optimization process with Taguchi DOE

Utilization of migratory locusts (Locusta migratoria) as a main substrate due to its high protein content for inulinase (2,1-β-d-fructan fructanohydrolase) production by Geotrichum candidum OC-7 was investigated in this study. To optimize fermentation conditions, four influential factors (locust powder (LP) concentration, sucrose concentration, pH and fermentation time) at three levels were investigated using Taguchi orthogonal array (OA) design of experiment (DOE). Inulinase yield obtained from the designed experiments with regard to Taguchi L9 OA was processed with Minitab 15 software at ‘larger is better’ as quality character. The results showed that optimal fermentation conditions determined as LP 30 g/l, sucrose 20 g/l, pH 6.0 and time 48 h. Maximum inulinase activity was recorded as 30.12 U/ml, which was closer to the predicted value (30.56 U/ml). To verify the results, analysis of variance test was employed. LP had the greatest contribution (71.96%) among the other factors. Sucrose had lower contribution (13.96%) than LP. This result demonstrated that LP had a strong effect on inulinase activity and can be used for enzyme production. Taguchi DOE application enhanced enzyme activity to about 3.05-fold versus unoptimized condition and 2.34-fold versus control medium. Consequently, higher inulinase production can be achieved by the utilization of an edible insect material as an alternative substrate and Taguchi DOE presents suitable optimization method for biotechnological process.