Search results for: phytase
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 6

Search results for: phytase

6 Cloning and Over Expression of an Aspergillus niger XP Phytase Gene (phyA) in Pichia pastoris

Authors: Ngo Thanh Xuan, Mai Thi Hang, Vu Nguyen Thanh

Abstract:

A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.

Keywords: Aspergillus niger XP, cloning, over expression, Pichia pastoris, phyA , phytase.

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5 Carbon Sources Utilization Profiles of Thermophilic Phytase Producing Bacteria Isolated from Hot-spring in Malaysia

Authors: Noor Muzamil Mohamad, Abdul Manaf Ali, Hamzah Mohd Salleh

Abstract:

Phytases (myo-inositol hexakisphosphate phosphohydrolases; EC 3.1.3.8) catalyze the hydrolysis of phytic acid (myoinositol hexakisphosphate) to the mono-, di-, tri-, tetra-, and pentaphosphates of myo-inositol and inorganic phosphate. Therrmophilic bacteria isolated from water sampled from hot spring. About 120 isolates of bacteria were successfully isolated form hot spring water sample and tested for extracellular phytase producing. After 5 passages of the screening on the PSM media, 4 isolates were found stable in producing phytase enzyme. The 16s RDNA sequencing for identification of bacteria using molecular technique revealed that all isolates those positive in phytase producing are belong to Geobacillus spp. And Anoxybacillus spp. Anoxybacillus rupiensis UniSZA-7 were identified for their carbon source utilization using Phenotype Microarray Plate of Biolog and found they utilize several kind of carbon source provided.

Keywords: Phytase, Phytic Acid, Thermophilic Bacteria, PSM Media and Phytase Assay

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4 Identification Characterization and Production of Phytase from Endophytic Fungi

Authors: Yetti Marlida , Rina Delfita , Neni Gusmanizar, Gita Ciptaan

Abstract:

Phytases are acid phosphatase enzymes, which efficiently cleave phosphate moieties from phytic acid, thereby generating myo-inositol and inorganic phosphate. Thirty four isolates of endophytic fungi to produce of phytases were isolated from leaf, stem and root fragments of soybean. Screening of 34 isolates of endophytic fungi identified the phytases produced by Rhizoctonia sp. and Fusarium verticillioides . The phytase production were the best induced by phytic acid and rice bran compared the others inducer in submerged fermentation medium used. The phytase produced by both Rhizoctonia sp. and F. verticillioides have pH optimum at 4.0 and 5.0 respectively. The characterization of phytase from Fusarium verticillioides showed that temperature optimum was 500C and stability until 600C, the pH optimum 5.0 and pH stability was 2.5 – 6.0, and substrate specificity were rice bran>soybean meal>corn> coconut cake, respectively.

Keywords: endophytic fungus, phytase, soybean, Rhizoctoniasp., Fusarium verticillioides,

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3 The Construction of a Probiotic Lactic Acid Bacterium Expressing Acid-Resistant Phytase Enzyme

Authors: R. Majidzadeh Heravi, M. Sankian, H. Kermanshahi, M. R. Nassiri, A. Heravi Moussavi, S. A. Lari, A. R. Varasteh

Abstract:

The use of probiotics engineered to express specific enzymes has been the subject of considerable attention in poultry industry because of increased nutrient availability and reduced cost of enzyme supplementation. Phytase enzyme is commonly added to poultry feed to improve digestibility and availability of phosphorus from plant sources. To construct a probiotic with potential of phytate degradation, phytase gene (appA) from E. coli was cloned and transformed into two probiotic bacteria Lactobacillus salivarius and Lactococcus lactis. L. salivarous showed plasmid instability, unable to express the gene. The expression of appA gene in L. lactis was analyzed by detecting specific RNA and zymography assay. Phytase enzyme was isolated from cellular extracts of recombinant L. lactis, showing a 46 kDa band upon the SDS-PAGE analysis. Zymogram also confirmed the phytase activity of the 46 kDa band corresponding to the enzyme. An enzyme activity of 4.9U/ml was obtained in cell extracts of L. lactis. The growth of native and recombinant L. lactis was similar in the presence of two concentrations of ox bile.

Keywords: Lactobacillus salivarus, Lactococcus lactis, recombinant, phytase, poultry.

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2 The Effects of Organic or Inorganic Zinc and Microbial Phytase, Alone or in Combination, on the Performance, Biochemical Parameters and Nutrient Utilization of Broilers Fed a Diet Low in Available Phosphorus

Authors: Mustafa Midilli, Mustafa Salman, Omer Hakan Muglali, Tülay Ögretmen, Sena Cenesiz, Neslihan Ormanci

Abstract:

This study examined the effects of zinc (Zn) from different sources and microbial phytase on the broiler performance, biochemical parameters and digestibility of nutrients when they were added to broiler diets containing low available phosphorus. A total of 875, 1-day-old male broilers of the Ross 308 strain were randomly separated into two control groups (positive and negative) and five treatment groups each containing 125 birds; each group was divided into 5 replicates of 25 birds. The positive control (PC) group was fed a diet containing adequate concentration (0.45%) of available phosphorus due to mineral premix (except zinc) and feeds. The negative control (NC) group was fed a basal diet including low concentration (0.30%) of available phosphorus due to mineral premix (except zinc) and feeds. The basal diet was supplemented with 0.30% phosphorus and 500 FTU phytase (PH); 0.30% phosphorus and organic zinc (OZ; 75mg/kg of Zn from Zn-proteinate); 0.30% phosphorus and inorganic zinc (IZ; 75 mg/kg of Zn from ZnSO4); 0.30% phosphorus, organic zinc and 500 FTU phytase (OZ + PH); and 0.30% phosphorus, inorganic zinc and 500 FTU phytase (IZ + PH) in the treatment groups 1, 2, 3, 4 and 5, respectively. The lowest value for mean body weight was in the negative control group on a diet containing low available phosphorus. The use of supplementation with organic and inorganic zinc alone or in combination with microbial phytase significantly (P<0.05) increased the digestibility of Zn in the male broilers. Supplementation of those diets with OZ + PH or IZ + PH was very effective for increasing the body weight, body weight gain and the feed conversion ratio. In conclusion, the effects on broilers of diets with low phosphorus levels may be overcome by the addition of inorganic or organic zinc compounds in combination with microbial phytase.

Keywords: Broiler, Performance, Phytase, Phosphorus, Zinc.

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1 Stability of New Macromycetes Phytases under Room, Cooling and Freezing Temperatures of Storage

Authors: Michele R. Spier, Denise N. X. Salmon, Renato L. Binati, Luíza C. Piva, Adriane B.P. Medeiros, Carlos R. Soccol

Abstract:

Phytases are enzymes used as an important component in monogastric animals feeds in order to improve phosphorous availability, since it is not readily assimilated by these animals in the form of the phytate presented in plants and grains. As these enzymes are used in industrial activities, they must retain its catalytic activities during a certain storage period. This study presents information about the stability of 4 different phytases, produced by four macromycetes fungi through solid-state fermentation (SSF). There is a lack of data in literature concerning phytase from macromycetes shelf-life in storage conditions at room, cooling and freezing temperatures. The 4 phytases from macromycetes still had enzymatic activities around 100 days of storage at room temperature. At cooling temperature in 146 days of studies, the phytase from G. stipitatum was the most stable with 44% of the initial activity, in U.gds (units per gram of dried fermented substrate). The freezing temperature was the best condition storage for phytases from G. stipitatum and T. versicolor. Each condition provided a study for each mushroom phytase, totalizing 12 studies. The phytases showed to be stable for a long period without the addition of additives.

Keywords: macromycetes, phytase, solid-state fermentation, wheat bran, stability

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