@article { ISI:000249921800011, title = {Production and partial characterization of two types of phytase from aspergillus niger NCIM 563 under submerged fermentation conditions}, journal = {World Journal of Microbiology \& Biotechnology}, volume = {23}, number = {11}, year = {2007}, month = {NOV}, pages = {1585-1593}, publisher = {SPRINGER}, type = {Article}, address = {233 SPRING ST, NEW YORK, NY 10013 USA}, abstract = {

Novel extracellular phytase was produced by Aspergillus niger NCIM 563 under submerged fermentation conditions at 30 degrees C in medium containing dextrin and glucose as carbon sources along with sodium nitrate as nitrogen source. Maximum phytase activity (41.47 IU/mL at pH 2.5 and 10.71 IU/mL at pH 4.0) was obtained when dextrin was used as carbon source along with glucose and sodium nitrate as nitrogen source. Nearly 13 times increase in phytase activity was observed when phosphate in the form of KH2PO4 (0.004 g/100 mL) was added in the fermentation medium. Physic-chemical properties of partially purified enzyme indicate the possibility of two distinct forms of phytases, Phy I and Phy II. Optimum pH and temperature for Phy I was 2.5 and 60 degrees C while Phy II was 4.0 and 60 degrees C, respectively. Phy I was stable in the pH range 1.5-3.5 while Phy II was stable in the wider pH range, 2.0-7.0. Molecular weight of Phy I and Phy II on Sephacryl S-200 was approximately 304 kDa and 183 kDa, respectively. Phy I activity was moderately stimulated in the presence of 1 mM Mg2+, Mn2+, Ca2+ and Fe3+ ions and inhibited by Zn2+ and Cd2+ ions while Phy II activity was moderately stimulated by Fe3+ ions and was inhibited by Hg2+, Mn2+ and Zn2+ ions at 1 mM concentration in reaction mixture. The Km for Phy I and II was 3.18 and 0.514 mM while Vmax was 331.16 and 59.47 mu mols/min/mg protein, respectively.

}, keywords = {Aspergillus niger, Phytase, phytate degradation, Poultry feed supplement, Submerged fermentation}, issn = {0959-3993}, doi = {10.1007/s11274-007-9404-9}, author = {Soni, Sarvesh Kumar and Khire, Jayant Malhar} } @article { ISI:000255865500030, title = {Influence of pretreatment of agriculture residues on phytase production by aspergillus niger NCIM 563 under submerged fermentation conditions}, journal = {African Journal of Biotechnology}, volume = {7}, number = {8}, year = {2008}, month = {APR}, pages = {1101-1106}, publisher = {ACADEMIC JOURNALS}, type = {Article}, address = {P O BOX 5170-00200 NAIROBI, VICTORIA ISLAND, LAGOS 73023, NIGERIA}, abstract = {

The extracellular phytase production by Aspergillus niger NCIM 563 was evaluated in medium containing various agriculture residues under submerged fermentation conditions. Phytase production was affected by inorganic phosphate content of agriculture residues which ranged from 2.8 to 8 mg/g. The agriculture residues containing less than 4 mg/g inorganic phosphate supported phytase production with maximum activity of 68 IU/ml in medium containing 1\% rice bran on 11(th) day of fermentation. Addition of glucose up to 5\% in fermentation medium containing 1\% rice bran, enhanced phytase production. Pretreatment of agriculture residues with water to remove excess inorganic phosphate has significantly enhanced the phytase activity in case of de-oiled rice bran, wheat bran, peanut cake (low and high oil) and coconut cake. Maximum increase of 20.3 times in phytase activity was observed in case of wheat bran as compared to de-oiled rice bran, coconut cake, peanut cake high and low oil in which the increase in phytase activity was 6.85, 6.1, 5.3 and 3.0 times, respectively. Maximum phytase activity of 68 IU/ml was produced on the 11(th) day of fermentation compared to earlier reported 41.47 IU/ml phytase activity on the 15(th) day of submerged fermentation using 5\% dextrin and 2.5\% glucose, thus increasing productivity.

}, keywords = {Agriculture residue, Aspergillus niger, Phytase, phytate degradation, Poultry feed supplement, Submerged fermentation}, issn = {1684-5315}, author = {Bhavsar, Kavita P. and Shah, Parin and Soni, Sarvesh Kumar and Khire, Jayant Malhar} } @article { ISI:000263683800007, title = {Strain improvement and up scaling of phytase production by aspergillus niger NCIM 563 under submerged fermentation conditions}, journal = {Journal of Industrial Microbiology \& Biotechnology}, volume = {36}, number = {3}, year = {2009}, month = {MAR}, pages = {373-380}, publisher = {SPRINGER HEIDELBERG}, address = {TIERGARTENSTRASSE 17, D-69121 HEIDELBERG, GERMANY}, abstract = {

Combination of physical and chemical mutagenesis was used to isolate hyper secretory strains of Aspergillus niger NCIM 563 for phytase production. Phytase activity of mutant N-1 and N-79 was about 17 and 47\% higher than the parent strain. In shake flask the productivity of phytase in parent, mutant N-1 and N-79 was 6,181, 7,619 and 9,523 IU/L per day, respectively. Up scaling of the fermentation from shake flask to 3 and 14 L New Brunswick fermenter was studied. After optimizing various fermentation parameters like aeration, agitation and carbon source in fermentation medium the fermentation time to achieve highest phytase activity was reduced considerably from 14 days in shake flask to 8 days in 14 L fermenter. Highest phytase activity of 80 IU/ml was obtained in 1\% rice bran-3.5\% glucose containing medium with aeration 0.2 vvm and agitation 550 rpm at room temperature on 8th day of fermentation. Addition of either bavistin (0.1\%), penicillin (0.1\%), formalin (0.2\%) and sodium chloride (10\%) in fermented broth were effective in retaining 100\% phytase activity for 8 days at room temperature while these reagents along with methanol (50\%) and ethanol (50\%) confer 100\% stability of phytase activity at 4A degrees C till 20 days. Among various carriers used for application of phytase in feed, wheat bran and rice bran were superior to silica and calcium carbonate. Thermo stabilization studies indicate 100\% protection of phytase activity in presence of 12\% skim milk at 70A degrees C, which will be useful for its spray drying.

}, keywords = {Agriculture residue, Aspergillus niger, Phytase, phytate degradation, Poultry feed supplement, Submerged fermentation}, issn = {1367-5435}, doi = {10.1007/s10295-008-0506-7}, author = {Shah, P. and Bhavsar, Kavita P. and Soni, S. K. and Khire, Jayant Malhar} } @article { ISI:000282507400010, title = {Purification and characterization of two distinct acidic phytases with broad pH stability from aspergillus niger NCIM 563}, journal = {World Journal of Microbiology \& Biotechnology}, volume = {26}, number = {11}, year = {2010}, month = {NOV}, pages = {2009-2018}, publisher = {SPRINGER}, address = {233 SPRING ST, NEW YORK, NY 10013 USA}, abstract = {

Aspergillus niger NCIM 563 produced two different extracellular phytases (Phy I and Phy II) under submerged fermentation conditions at 30A degrees C in medium containing dextrin-glucose-sodium nitrate-salts. Both the enzymes were purified to homogeneity using Rotavapor concentration, Phenyl-Sepharose column chromatography and Sephacryl S-200 gel filtration. The molecular mass of Phy I and II as determined by SDS-PAGE and gel filtration were 66, 264, 150 and 148 kDa respectively, indicating that Phy I consists of four identical subunits and Phy II is a monomer. The pI values of Phy I and II were 3.55 and 3.91, respectively. Phy I was highly acidic with optimum pH of 2.5 and was stable over a broad pH range (1.5-9.0) while Phy II showed a pH optimum of 5.0 with stability in the range of pH 3.5-9.0. Phy I exhibited very broad substrate specificity while Phy II was more specific for sodium phytate. Similarly Phy II was strongly inhibited by Ag(+), Hg(2+) (1 mM) metal ions and Phy I was partially inhibited. Peptide analysis by Mass Spectrometry (MS) MALDI-TOF also indicated that both the proteins were totally different. The K (m) for Phy I and II for sodium phytate was 2.01 and 0.145 mM while V (max) was 5,018 and 1,671 mu mol min(-1) mg(-1), respectively. The N-terminal amino acid sequences of Phy I and Phy II were FSYGAAIPQQ and GVDERFPYTG, respectively. Phy II showed no homology with Phy I and any other known phytases from the literature suggesting its unique nature. This, according to us, is the first report of two distinct novel phytases from Aspergillus niger.

}, keywords = {Aspergillus niger, Phytase, Phytate, Poultry feed, Purification}, issn = {0959-3993}, doi = {10.1007/s11274-010-0385-8}, author = {Soni, S. K. and Magdum, A. and Khire, Jayant Malhar} } @article { ISI:000294225100030, title = {High level phytase production by aspergillus niger NCIM 563 in solid state culture: response surface optimization, up-scaling, and its partial characterization}, journal = {Journal of Industrial Microbiology \& Biotechnology}, volume = {38}, number = {9}, year = {2011}, month = {SEP}, pages = {1407-1417}, publisher = {SPRINGER HEIDELBERG}, address = {TIERGARTENSTRASSE 17, D-69121 HEIDELBERG, GERMANY}, abstract = {

Phytase production by Aspergillus niger NCIM 563 was optimized by using wheat bran in solid state fermentation (SSF). An integrated statistical optimization approach involving the combination of Placket-Burman design (PBD) and Box-Behnken design (BBD) was employed. PBD was used to evaluate the effect of 11 variables related to phytase production, and five statistically significant variables, namely, glucose, dextrin, NaNO(3), distilled water, and MgSO(4)center dot 7H(2)O, were selected for further optimization studies. The levels of five variables for maximum phytase production were determined by a BBD. Phytase production improved from 50 IU/g dry moldy bran (DMB) to 154 IU/g DMB indicating 3.08-fold increase after optimization. A simultaneous reduction in fermentation time from 7 to 4 days shows a high productivity of 38,500 IU/kg/day. Scaling up the process in trays gave reproducible phytase production overcoming industrial constraints of practicability and economics. The culture extract also had 133.2, 41.58, and 310.34 IU/g DMB of xylanase, cellulase, and amylase activities, respectively. The partially purified phytase was optimally active at 55A degrees C and pH 6.0. The enzyme retained ca. 75\% activity over a wide pH range 2.0-9.5. It also released more inorganic phosphorus from soybean meal in a broad pH range from 2.5 to 6.5 under emulated gastric conditions. Molecular weight of phytase on Sephacryl S-200 was approximately 87 kDa. The K (m) and V (max) observed were 0.156 mM and 220 mu m/min/mg. The SSF phytase from A. niger NCIM 563 offers an economical production capability and its wide pH stability shows its suitability for use in poultry feed.

}, keywords = {Aspergillus niger, Phytase, Response surface optimization, Solid state fermentation, Statistical methods}, issn = {1367-5435}, doi = {10.1007/s10295-010-0926-z}, author = {Bhavsar, Kavita P. and Kumar, V. Ravi and Khire, Jayant Malhar} } @article { ISI:000309333600007, title = {Downstream processing of extracellular phytase from aspergillus niger: chromatography process vs. aqueous two phase extraction for its simultaneous partitioning and purification}, journal = {Process Biochemistry}, volume = {47}, number = {7}, year = {2012}, month = {JUL}, pages = {1066-1072}, publisher = {ELSEVIER SCI LTD}, address = {THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND}, abstract = {

The application of single step aqueous two-phase extraction (ATPE) for the downstream processing of phytase from Aspergillus niger NCIM 563, produced under solid state fermentation, has been studied and compared with the traditional multi-step procedure involving salt precipitation and column chromatography. High phytase recovery (98.5\%) within a short time (3 h) and improved thermostability was attained by ATPE in comparison to 20\% recovery in 96 h by chromatography process.The ATPE method, therefore, seems to be an interesting alternative for simultaneous partitioning and purification of phytase. The influence of system parameters; such as, phase forming salts, polymer molecular weight and system pH on the partitioning behavior of phytase was evaluated. The ATPE system consisting of combination of polyethylene glycol (PEG) 6000 and 8000(10.5\%) and sodium citrate (20.5\%) resulted in one-sided partitioning of phytase in bottom phase with a purification factor of 2.5. This is the first report on phytase purification using liquid-liquid extraction and the results are likely to be beneficial in the poultry feed industry. (c) 2012 Elsevier Ltd. All rights reserved.

}, keywords = {Aspergillus niger, liquid-liquid extraction, Phytase, Polyethylene glycol, Purification}, issn = {1359-5113}, doi = {10.1016/j.procbio.2012.03.012}, author = {Bhavsar, Kavita P. and Kumar, V. Ravi and Khire, Jayant Malhar} } @article { ISI:000313651700018, title = {Combinatorial approach of statistical optimization and mutagenesis for improved production of acidic phytase by Aspergillus niger NCIM 563 under submerged fermentation condition}, journal = {Applied Microbiology and Biotechnology}, volume = {97}, number = {2}, year = {2013}, month = {JAN}, pages = {673-679}, publisher = {SPRINGER}, address = {233 SPRING ST, NEW YORK, NY 10013 USA}, abstract = {

Combination of statistical optimization and mutagenesis to isolate hypersecretory strains is studied to maximize phytase production from Aspergillus niger NCIM 563 under submerged fermentation. The overall results obtained show a remarkable 5.98-fold improvement in phytase production rates when compared to that using basal medium. Optimization of culture conditions from parent strain is studied first by the Plackett-Burman technique to evaluate the effects of 11 variables for phytase production. The results showed that glucose, MgSO4, KCl, incubation period, and MnSO4 are the most significant variables affecting enzyme production. Further optimization in these variables, using a central composite design technique, resulted in 3.74-fold increase in the yield of phytase production to 254,500 U/l when compared with the activity observed with basal media (68,000 U/l) in shake flask. Our experiments show that the phytase from A. niger NCIM 563 exhibits desirable activity in simulated gastric fluid conditions with low pH and also improved thermostability when compared to commercial phytase. The improved yield demonstrates the potential applicability of phytase enzyme as a source of phytase supplement for phosphorus nutrition and environmental protection in animal feed industry. Physical and chemical mutagenesis experiments were carried out in parallel to isolate hypersecretory mutants that could possibly further enhance the enzyme production. Using optimized media conditions of the parent strain, our results show that mutant strain A. niger NCIM 1359 increased the phytase activity by another 1.6-fold to 407,200 U/l.

}, keywords = {Animal feed, Aspergillus niger, Mutagenesis, Phytase, Statistical optimization}, issn = {0175-7598}, doi = {10.1007/s00253-012-3965-8}, author = {Bhavsar, Kavita P. and Gujar, P. and Shah, P. and Kumar, V. Ravi and Khire, Jayant Malhar} } @article { ISI:000319875500022, title = {Effect of phytase from aspergillus niger on plant growth and mineral assimilation in wheat (Triticum aestivum Linn.) and its potential for use as a soil amendment}, journal = {Journal of the Science of Food and Agriculture}, volume = {93}, number = {9}, year = {2013}, month = {JUL}, pages = {2242-2247}, publisher = {WILEY-BLACKWELL}, address = {111 RIVER ST, HOBOKEN 07030-5774, NJ USA}, abstract = {

Background Extensive use of non-replenishable phosphate reserves as phosphate supplements in agriculture and animal feed poses a threat for environmental pollution and necessitated a search for alternative phosphate sources. Unlocking the phytate phosphorus using microbial phytase can provide an ecofriendly solution in agriculture. The aim of the present work was to evaluate the effect of phytase from Aspergillus niger NCIM 563 in degradation of phytate phosphorus to benefit plant nutrition and soil amendment in comparison with chemical fertilizers. Results An enzyme dose of 12 IU was sufficient to hydrolyze phytate and increase assimilation of phosphorus by about 74\%. Phytase supplementation leads to increase in shoot:total length ratio by about 200\%, indicating its growth-promoting effect. Consistency in phytase-induced growth was reflected at pot and tray levels, wherein shoot:total length ratio was observed to be 2.01 and 2.12 respectively. Mineral assimilation due to phytase was more efficient as compared to chemical fertilizers, thus overcoming the constraints of practicability and economics in the agriculture industry. Phytase was efficient in reducing the phytic acid content of soil by about 30\% while simultaneously increasing the phytate phosphate availability by 1.18-fold. Conclusion Phytase from A. niger showed improvement in phytate phosphorus and mineral availability. Besides a plant growth-promoting effect, reduction in use of chemical fertilizers and soil improvement could be achieved simultaneously for maintaining the sustainability of agriculture. (c) 2012 Society of Chemical Industry

}, keywords = {mineral assimilation, Phytase, phytate degradation, plant growth promotion, soil amendment}, issn = {0022-5142}, doi = {10.1002/jsfa.6032}, author = {Gujar, Pradnya D. and Bhavsar, Kavita P. and Khire, Jayant Malhar} } @article { ISI:000327920600003, title = {Phytase isozymes from aspergillus niger NCIM 563 under solid state fermentation: biochemical characterization and their correlation with submerged phytases}, journal = {Process Biochemistry}, volume = {48}, number = {11}, year = {2013}, month = {NOV}, pages = {1618-1625}, publisher = {ELSEVIER SCI LTD}, address = {THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND}, abstract = {

Aspergillus niger NCIM 563 produces dissimilar phytase isozymes under solid state and submerged fermentation conditions. Biochemical characterization and applications of phytase Phy III and Phy IV in SSF and their comparison with submerged fermentation Phy I and Phy III were studied. SSF phytases have a higher metabolic potential as compared to SmF. Phy I is tetramer and Phy II, III and IV are monomers. Phy I and IV have pH optima of 2.5 and Phy II and III have pH optima of 5.0 and 5.6, respectively. Phy I, III and IV exhibited very broad substrate specificity while Phy II was more specific for sodium phytate. SSF phytase is less thermostable as compared to SmF phytase. Phy land II show homology with other known phytases while Phy III and IV show no homology with SmF phytases and any other known phytases from the literature suggesting their unique nature. This is the first report about differences among phytase produced under SSF and SmF by A. niger and this study provides basis for explanation of the stability and catalytic differences observed for these enzymes. Exclusive biochemical characteristics and multilevel application of SSF native phytases determine their efficacy and is exceptional. (C) 2013 Elsevier Ltd. All rights reserved.

}, keywords = {A. niger, Isozymes, Phytase, Solid state fermentation, Submerged fermentation}, issn = {1359-5113}, doi = {10.1016/j.procbio.2013.08.001}, author = {Bhavsar, Kavita P. and Buddhiwant, Priyanka and Soni, Sarvesh K. and Depan, Dilip and Sarkar, Sampa and Khire, Jayant Malhar} } @article {46098, title = {Use of Plackett-Burman design for enhanced phytase production by Williopsis saturnus NCIM 3298 for applications in animal feed and ethanol production}, journal = {3 Biotech}, volume = {9}, year = {2019}, month = {JUN}, pages = {237}, type = {Article}, abstract = {

Distiller-dried grain solid (DDGS), a co-product of alcohol production, contains cereal grain residues, proteins, and yeast metabolites, which make it suitable in poultry feeding. However, high phytate content of DDGS limits its applicability in poultry feed. In this study, Plackett-Burman design was used to improve cell-bound phytase production by Williopsis saturnus NCIM 3298, and we achieved an enzyme activity of 269IU/g of dry-wet biomass. The effect of this enhanced phytase-displaying yeast strain on hydrolysis of corn phytate and subsequently on ethanol production and DDGS quality was then investigated. Results of saccharification in the presence of phytase showed that reducing sugar content of liquefied mash increased by 11\%, which subsequently improved the ethanol production by 18\% (w/v) (p\<0.01) compared with the control. Notably, phytase treatment decreased the phytate content of corn by 70\% (p\<0.01) compared with the control, thereby improving the availability of free phosphate in fermentation broth and DDGS. Thus, the results obtained suggest that the addition of W. saturnus NCIM 3298 strain has the potential of providing a new source of phytase that would be useful in the feed and ethanol industries.

}, keywords = {DDGS, Ethanol, Phytase, Plackett-Burman Design, Williopsis saturnus NCIM 3298}, issn = {2190-572X}, doi = {10.1007/s13205-019-1764-y}, author = {Pable, Anupama A. and Shah, Sarah and Kumar, V. Ravi and Khire, Jayant M.} } @article {47778, title = {Performance of Aspergillus niger (NCIM 563) phytase based feed supplement for broiler growth and phosphorus excretion}, journal = {Biocatalysis and Agricultural Biotechnology}, volume = {31}, year = {2021}, month = {JAN}, pages = {101887}, type = {Article}, abstract = {Despite availability of commercial enzymes, the phytase produced from relatively inexpensive systems with high yields are gaining global attention in the feed industries in post-antibiotic era. We studied A. niger NCIM 563 Phytase produced in solid state fermentation (SSF) derived Koji powder and evaluated its utility in the poultry feed for broiler growth performance and phosphorous (P) excretion. The ability of phytase in the dried powder was estimated to dephytinize the poultry feed under simulated gastric conditions. Poultry feed was formulated using A. niger NCIM 563 phytase followed by a 42 days feed trial on broilers. After supplementation of phytase to the diet, there was a reduction of dietary P, maintained growth performance, skeletal development of broilers and reduced levels of phytic acid and available P in the litter. Extracellular phytase was able to replace up to 0.1\% P in poultry feed. Minimal downstream processing a low-cost feed supplement with significant phytase activity could provide added advantage for anti-nutrition free poultry feed.}, keywords = {Phosphorus, Phytase, Poultry feed, Solid state fermentation}, isbn = {1878-8181}, doi = {10.1016/j.bcab.2020.101887}, url = {https://www.sciencedirect.com/science/article/pii/S1878818120319186}, author = {Puppala, Kumar Raja and Buddhiwant, Priyanka G. and Agawane, Sachin B. and Kadam, Avinash S. and Mote, Chandrashekhar S. and Lonkar, Vijaysinh D. and Khire, Jayant M. and Dharne, Mahesh S.} } @article {48464, title = {Isolation and biochemical characterization of acid phytase from aspergillus niger and its applications in dephytinization of phytic acid in poultry feed ingredients}, journal = {Microbiology}, volume = {92}, year = {2023}, month = {APR}, pages = {221-229}, type = {Article}, abstract = {

Acidic phytase (PHY-B) was isolated from Aspergillus niger BIONCL8 strain, and assessed its application for the dephytinization of poultry feed ingredients. Aspergillus niger BIONCL8 strain was identified by precise molecular methods by targeting internal transcribed spacer (ITS) region of the fungi and molecular detection of PHY gene in the strain was confirmed by sequence analysis with respect to PHY production. The enzyme was isolated and purified by DEAE Sephadex A-50 and Bio-Gel P-60 Gel ion-exchange chromatography. The estimated molecular weight of the protein was 65 kDa on SDS-PAGE, it had a specific activity of 21.18 U/mg. The molecular weight of the protein was confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The peptide sequence of PHY-B was identified using high resolution-mass spectrometry (LC-HR-MS), and its physicochemical parameters were studied. The PHY was stable at acidic pH 2.1, most active at 40 degrees C and was stable up to 80 degrees C, retaining 30\% residual activity after 1 h of incubation. The PHY activity was enhanced in the presence of Mg2+ and EDTA, and activity was inhibited in the presence of Hg2+ and K+. The K-m and V-max of PHY were recorded as 3.35 mM and 1.27 U/mg, respectively. The PHY was shown to play a significant role in decreasing PA content in various poultry feed ingredients, ranging from 48.14 to 82.14\%. The novel features of the enzyme can be used to decrease the PA content in feed ingredients and increase the bioavailability of nutrients to non-ruminant animals.

}, keywords = {Aspergillus niger, dephytinization, high resolution-mass spectrometry, ion-exchange chromatography, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, Phytase}, issn = {0026-2617}, doi = {10.1134/S0026261722601452}, author = {Bhandari, Y. and Sonwane, B. and Vamkudoth, K. R.} }