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Production and characterization of pullulan from beet molasses using a nonpigmented strain of Aureobasidium pullulans in batch culture.

The production of pullulan from beet molasses by a pigment-free strain of Aztreobasidium pullulans on shake-flask culture was investigated. Combined pretreatment of molasses with sulfuric acid and activated carbon to remove potential fermentation inhibitors present in molasses resulted in a maximum pullulan concentration of 24 g/L, a biomass dry wt of 14 g/L, a pullulan yield of 52.5%, and a sugar utilization of 92% with optimum fermentation conditions (initial sugar concentration of 50 g/L and initial pH of 7.0). The addition of other nutrients as carbon and nitrogen supplements (olive oil, ammonium sulfate, yeast extract) did not further improve the production of the exopolysaccharides. Structural characterization of the isolated polysaccharides from the fermentation broths by 13C-nuclear magnetic resonance spectroscopy and pullulanase digestion combined with size-exclusion chromatography confirmed the identity of pullulan and the homogeneity (>93% dry basis) of the elaborated polysaccharides by the microorganism. Using multiangle laser light scattering and refractive index detectors in conjunction with high-performance size-exclusion chromatography molecular size distributions and estimates of the molecular weight (Mw = 2.1-4.1 x 10(5)), root mean square of the radius of gyration (R = 30-38 nm), and polydispersity index (Mw/Mn = 1.4-2.4) were obtained. The fermentation products of molasses pretreated with sulfuric acid and/or activated carbon were more homogeneous and free of contaminating proteins. In the concentration range of 2.8-10.0 (w/v), the solution's rheologic behavior of the isolated pullulans was almost Newtonian (within 1 and 1200 s(-1) at 20 degrees C); a slight shear thinning was observed at 10.0 (w/v) for the high molecular weight samples. Overall, beet molasses pretreated with sulfuric acid and activated carbon appears as an attractive fermentation medium for the production of pullulan by A. pullulans.

Ascomycota↗

Value of a molasses mixture containing natural protein as a supplement to brood cows offered low-quality forages.

In a 4-yr study, molasses (M; 6% CP), molasses-urea (MU; 17.5% CP; 92% molasses, 4% urea and 4% water) and molasses-cottonseed meal-urea (MCSM; 17.5% CP; 73% molasses, 25% cottonseed meal, 1% urea and 1% water) were compared as winter supplements for Braford cows (514 matings) consuming low-quality forage. Cows were fed 1.35, 1.50 and 1.27 kg.head-1.d-1 of the respective supplements for 127 d beginning in December. Cows were wintered on bahiagrass pasture (avg 8% CP) and stargrass hay (avg 5% CP). Quantity of hay offered averaged 7.6 kg.cow-1.d-1 over 109 d. Cows were exposed to bulls for 90 d beginning on March 1. Supplementation treatment did not affect cow live weight, live weight change or condition score (P greater than .05). However, pregnancy rate was higher (P less than .05) for cows fed MCSM (79.0%) than for cows fed M (67.8%). The pregnancy rate of cows fed MU (76.3%) was intermediate to, but not different (P greater than .05) from, that of cows fed either M or MCSM. Supplementation treatment did not affect calf performance as measured by birth weight, survival rate or weaning weight (P greater than .05). However, calves from cows fed MCSM were 6 d older (P less than .05) at weaning than calves from cows fed M. For 3-yr-old cows, pregnancy rate was considerably higher (P less than .05) for cows fed MCSM (69.6%) than for cows fed M (37.5%).(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Intake and digestion by Holstein steers consuming diets based on litter harvested after different numbers of broiler growing periods or with molasses addition before deep-stacking.

We determined the effects on feed intake and digestibility by Holstein steers of 1) the number of broiler growing periods before litter harvest and dietary level of broiler litter and 2) level of molasses added to broiler litter before deep-stacking. In Exp. 1, eight steers (179+/-7.4 kg average BW) were used in two simultaneous 4 x 4 Latin squares (2 x 4 factorial) with 21-d periods. Broiler litter harvested after one, three, or six 6-wk growing periods (1P, 3P, and 6P, respectively) mixed with .5 or 1.5% BW of ground corn (.5C and 1.5C, respectively) was consumed ad libitum. Bermudagrass hay was fed to 1P, 3P, and 6P steers at .5% BW and was ingested ad libitum by Control steers, along with feeding of .5 or 1.5% BW of corn (DM basis). Broiler litter was 63, 43, and 35% NDF, 2.2, 3.5, and 4.1% N, and 18, 30, and 27% ash for 1P, 3P, and 6P, respectively. Total tract digestibility of NDF was 53.7, 29.4, 50.4, 58.1, 31.3, 30.8, 34.1, and 49.5% (SE 3.50), and digestible OM intake was 2.21, 1.70, 2.27, 2.39, 2.26, 3.18, 2.93, and 3.34 kg/d (SE .160) for .5C-Control, .5C-1P, .5C-3P, .5C-6P, 1.5C-Control, 1.5C-1P, 1.5C-3P, and 1.5C-6P, respectively. In Exp. 2, five steers (228+/-6.0 kg average BW) were used in a 5 x 5 Latin square with 21-d periods. Offered diets were 15% bermudagrass hay and 60% broiler litter (6P of Exp. 1; DM). Molasses was offered at 0, 3.2, or 6.7% of total DM, with the balance of the diet composed of corn. Molasses for two treatments was mixed with litter at meals, whereas for two other treatments molasses and litter were mixed before deep-stacking. Only a few minor treatment effects on intakes and digestibilities occurred. In conclusion, digestible OM intake by growing steers was less for litter harvested after one broiler growing period than after three or six when fed with only .5% BW of corn, although the effect of the number of periods was negligible with corn given at 1.5% BW. Molasses addition before deep-stacking or at meals did not enhance feeding value of litter harvested after six broiler growing periods.

Animal Feed↗

Effect of corn- vs molasses-based supplements on trace mineral status in beef heifers.

Two studies were conducted to compare the availability of trace minerals offered to Brahman-crossbred heifers in either grain- or molasses-based supplements. Heifers were randomly assigned to bahiagrass pastures of equal size (n = 3 and 2 heifers/ pasture with 6 and 4 pastures/treatment for Experiment 1 and 2, respectively). Two supplements were formulated using corn and cottonseed meal (DRY) or molasses and cottonseed meal (LIQ). In Experiment 2, a third treatment consisted of the DRY supplement with additional S to equal the amount naturally supplied by the LIQ treatment (DRY+S). Supplements were formulated to provide, on average, 1.5 kg of TDN and 0.3 kg of CP/heifer daily and were fed three times weekly. Supplements also were fortified to provide 140, 76, and 63 mg of Cu, Mn, and Zn per heifer daily. Individual heifer weights were collected at the start and conclusion of the study, following a 12-h shrink. Plasma ceruloplasmin and liver Cu, Mn, Mo, Fe, and Zn concentrations were determined on d 0, 29, 56, and 84 in Experiment 1, and d 0, 32, 57, and 90 in Experiment 2. No differences were detected in heifer BW change (-9.3 and -7.3 kg for DRY and LIQ in Experiment 1, and 51.7, 46.3, and 46.7 kg for DRY, DRY+S, and LIQ in Experiment 2, respectively). In both experiments, liver Fe, Mn, and Zn concentrations were not affected by supplement treatment. Molybdenum tended (P = 0.06 and 0.10 for Experiments 1 and 2, respectively) to accumulate in the liver of heifers fed molasses-based supplements. In Experiment 1, Cu accumulation was less (P < 0.001) in heifers fed the liquid supplements (271 vs 224, 286 vs 202, and 330 vs 218 ppm, for DRY and LIQ supplements on d 29, 56, and 84, respectively). In Experiment 2, heifers receiving Cu from DRY supplements had a 155-ppm increase in liver Cu concentration, which was greater (P = 0.03) than DRY+S (87 ppm increase) and LIQ (P < 0.001; 13 ppm increase). Although lower than heifers receiving DRY, heifers receiving DRY+S had greater (P = 0.02) liver Cu concentrations than heifers receiving LIQ by the end of the study. In both experiments, plasma ceruloplasmin concentrations were highest (P < 0.04) in heifers receiving DRY supplement. The results of these studies suggest that components in molasses-based supplements decrease the accumulation of Cu in the liver of beef heifers. The S and Mo components of molasses may be responsible, at least in part, for this antagonism.

Animal Feed↗

Effect of copper source and level on performance and copper status of cattle consuming molasses-based supplements.

Two studies were conducted to evaluate the availability of dietary Cu offered to growing beef cattle consuming molasses-based supplements. In Exp. 1, 24 Braford heifers were assigned randomly to bahiagrass (Paspalum notatum) pastures (two heifers/pasture). Heifers were provided 1.5 kg of TDN and 0.3 kg of supplemental CP/heifer daily using a molasses-cottonseed meal slurry. Three treatments were randomly assigned to pastures (four pastures/treatment), providing 100 mg of supplemental Cu daily in the form of either CuSO4 (inorganic Cu) or organic-Cu. A third treatment offered no supplemental Cu (negative control). Heifer BW was collected at the start and end of the study. Jugular blood and liver samples were collected on d 0, 29, 56, and 84. In Exp. 2, 24 Brahman-crossbred steers were fed the same molasses-cottonseed meal supplement at the same rates used in Exp. 1. Steers were housed in individual pens (15 m2) with free-choice access to stargrass (Cynodon spp.) hay. Four Cu treatments were assigned to individual steers (six pens/treatment) providing 1) 10 ppm of Cu from an organic source; 2) 10 ppm Cu from Tri-basic Cu chloride (TBCC); 3) 30 ppm of Cu from TBCC; or 4) 30 ppm of Cu, a 50:50 ratio of TBCC and organic Cu. Body weights and jugular blood and liver samples were collected on d 0, 24, 48, and 72. In Exp. 1, liver Cu concentrations did not differ between heifers supplemented with inorganic and organic Cu. Each source resulted in increased (P < 0.05) liver Cu concentrations compared with the unsupplemented control. Plasma ceruloplasmin concentrations were higher (P < 0.05) for Cu-supplemented heifers, independent of Cu source. Heifer ADG tended (P = 0.11) to increase with Cu supplementation compared with the unsupplemented control. In Exp. 2, liver Cu was greater (P < 0.05) on d 24, 48, and 72 for steers consuming 30 vs. 10 ppm of Cu. Steers supplemented with organic Cu had lower DMI than steers supplemented with 10 or 30 ppm of TBCC. These data suggest that the inorganic and organic Cu sources evaluated in these studies were of similar availability when offered in molasses supplements. A dietary Cu concentration greater than 10 ppm might be necessary to ensure absorption in beef cattle fed molasses-based supplements.

Animal Feed↗

Effect of feeding urea-molasses blocks with incorporated fenbendazole on grazing dairy heifers naturally infected with gastrointestinal nematodes.

Between June 1999 and August 2000, the effects of feeding medicated urea-molasses supplement blocks on the growth of dairy heifers in a marginal area of central Kenya were assessed by comparing the live-weight gain of supplemented and unsupplemented heifers grazing the same pasture. Thirty-nine heifers with an average age of 9.6 months were initially treated orally with albendazole (10 mg/kg body weight) and assigned to 3 groups: group I was fed urea-molasses blocks with incorporated fenbendazole (MUMB), group II was fed urea-molasses blocks (UMB) and group III heifers (control) received no block supplementation (NBS). Body weights of the heifers and faecal egg counts (FECs) were measured monthly and larval cultures were made of positive faecal samples of each group. The mean cumulative live-weight responses of the MUMB and UMB groups were significantly greater than the NBS group (P < 0.05). However, at the end of the experimental period, the mean weight gain of the MUMB group did not differ from that of the UMB group (P > 0.05). The FECs were moderate to low in all groups and decreased progressively with increasing age of the animals; FECs for the urea-molasses-supplemented groups remained significantly lower than those of the NBS group throughout the experimental period (P < 0.05). Haemonchus and Trichostrongylus were the predominant nematode genera found in the heifers, but Cooperia, Bunostomum and Oesophagostomum were also present. These results indicate that feeding of urea-molasses blocks substantially reduced production losses attributable to nematode infection of young grazing cattle, and confirms previous observations that well-fed animals are better able to overcome the effects of helminth infections.

Administration, Oral↗

[Effects of sugar cane molasses on the nutritive value of Canavalia ensiformis seeds for broiler chicks].

Three experiments were conducted to determine the efficacy of sugar cane molasses to improve performance of broiler chicks fed a diet containing 30% raw or autoclaved Canavalia ensiformis seeds (Jack beans). For this latter purpose, canavalia seeds were ground and autoclaved at 120 degrees C and 15 psi during 60 min. Day-old male chicks (Cobb x Cobb) were used throughout the study. In Experiment 1, adding 10% molasses to a control diet devoided of jack beans seeds significantly (p < 0.05) increased chick feed intake. However, neither the addition of 10% molasses nor of 5% glucose, sucrose, fructose, xylose or corn starch to the 30% raw canavalia ration allowed feed intakes similar to that shown by the control diet (Experiment 2). Broiler performance was evaluated in Experiment 3 in response to diets containing 30% raw or autoclaved jack bean meal. Ten-percent molasses was also added to both diets which along with the control diet were fed to chiks had free access to diets. Growth was depressed when the Jack bean containing diets were pair-fed to chicks. The results indicated that the use of molasses does not overcome the deleterious effects on chick performance due to the presence of 30% raw or autoclaved jack bean meal in the diets.

Animal Feed↗

Utilization of beet molasses for riboflavin production by Mycobacterium phlei.

Mycobacterium phlei was tested for its ability to utilize beet molasses as the sole carbon source and produce riboflavin. The crude beet molasses was analyzed and treated in various ways to reduce its heavy element content and to remove the muddy residue. Promising amounts of riboflavin were produced when the organism was cultivated on decationized (resin-treated) beet molasses. The highest vitamin productivity was achieved by incubating the inoculated medium containing 9% molasses and initially adjusted to pH 6 under shacked condition for 6 days in the dark.

Fermentation↗

Statistical optimization of medium components for enhanced acetoin production from molasses and soybean meal hydrolysate.

The nutritional requirements for acetoin production by Bacillus subtilis CICC 10025 were optimized statistically in shake flask experiments using indigenous agroindustrial by-products. The medium components considered for initial screening in a Plackett-Burman design comprised a-molasses (molasses submitted to acidification pretreatment), soybean meal hydrolysate (SMH), KH(2)PO(4).3H(2)O, sodium acetate, MgSO(4).7H(2)O, FeCl(2), and MnCl(2), in which the first two were identified as significantly (at the 99% significant level) influencing acetoin production. Response surface methodology was applied to determine the mutual interactions between these two components and optimal levels for acetoin production. In flask fermentations, 37.9 g l(-1) acetoin was repeatedly achieved using the optimized concentrations of a-molasses and SMH [22.0% (v/v) and 27.8% (v/v), respectively]. a-Molasses and SMH were demonstrated to be more productive than pure sucrose and yeast extract plus peptone, respectively, in acetoin fermentation. In a 5-l fermenter, 35.4 g l(-1) of acetoin could be obtained after 56.4 h of cultivation. To our knowledge, these results, i.e., acetoin yields in flask or fermenter fermentations, were new records on acetoin fermentation by B. subtilis.

Acetoin↗

Biosynthesis of medium-chain-length poly(hydroxyalkanoates) from soy molasses.

Pseudomonas corrugata was selected from a screening process for the bioconversion of inexpensive soy molasses into medium-chain-length poly(hydroxyalkanoates) (mcl-PHA). We obtained yields of 1.5 g cell dry weight (CDW)/l culture with growth medium supplemented with 2% (w/v) soy molasses, and of an average of 3.4 g CDW/l with 5% (w/v) soy molasses. Crude PHAs were obtained at 5-17% of CDW. The most prominent repeat-unit monomers in the PHAs were 3-hydroxydodecanoate, 3-hydroxyoctanoate, 3-hydroxydodecanoate, and 3-hydroxytetradecenoate. This work represents the first description of fermentative mcl-PHA production from the soy molasses.

Caprylates↗

Optimization of nutrients for gellan gum production by Sphingomonas paucimobilis ATCC-31461 in molasses based medium using response surface methodology.

A molasses based medium for the production of gellan by Sphingomonas paucimobilis ATCC-31461 was developed. Placket-Burman design criterion was applied to study the effect of various nutrient supplements on gellan production using molasses. Among the 20 variables tested, molasses, tryptone, casaminoacid, disodium hydrogen orthophosphate and manganese chloride showed significant effect on gellan production. A central composite design was applied to determine the optimum concentrations of the significant variables obtained from Placket-Burman design. Most suitable medium composition for production of gellan was (g/l): molasses-112.5; tryptone-1; casaminoacid-1; disodium hydrogen orthophosphate-1; manganese chloride-0.947 and the optimum gellan production was 13.814 g/l.

Culture Media↗

Determination of sugars (and betaine) in molasses by high-performance liquid chromatography. Comparison of the results with those obtained by the classical Lane-Eynon method.

The adaptability of four different high-performance liquid chromatographic (HPLC) columns (ion exchangers in the sodium and calcium forms, ODS and NH2) for the analysis of sugars in molasses was studied. Some sample pre-treatment procedures were also examined. A strong cation-exchange column in the sodium form was the most suitable for the determination of both sugars in cane molasses and sugars and betaine in beet molasses. The only necessary pre-treatment was filtration of the diluted molasses sample through a 0.22-micron membrane before injection. HPLC gave lower sugar contents than the classical Lane-Eynon method. The lower values are more reliable, because the HPLC method eliminates interferents from the sugars.

Betaine↗

Enzymatic hydrolysis of molasses.

Kinetic studies of the enzymatic hydrolysis of molasses were conducted using glucoamylase. Central Sugar Refinery SDN BHD contains 13-20% glucose. The molasses was diluted and the kinetic experiments were conducted at 67 degrees C with 100-1000 mg/l of glucoamylase. The glucose contents of the molasses were enhanced after hydrolysis of molasses solution with 1000 mg/l glucoamylase. A Lineweaver-Burk plot was obtained based on enzyme kinetic data. The rate constant, Km and maximum reaction rate, Vmax for 500 mg/l of glucoamylase were 100 mmol/l (18 g/l) and 5 mmol/l min (0.9 g/l min), respectively. The maximum reaction rate, Vmax for 1000 mg/l of glucoamylase was doubled, to 100 mmol/l (18 g/l) and the rate constant, Km was the same for 500 mg/l of glucoamylase. The substrate inhibition model was noncompetitive based on the resulting Lineweaver-Burk plot for enzyme concentration of 500 and 1000 mg/l.

Color↗

Assessment of molasses-urea blocks for goat and sheep production in the Sultanate of Oman: intake and growth studies.

The goal of this study was to develop a molasses-urea block (MUB) for purposes of supplementing trace minerals to domestic ruminant livestock in Oman. To accomplish this, the utility of molasses and date syrup as fermentable energy sources, of straw, date flakes and wheat bran as fibre sources, and of cement and lime as binders were evaluated. The proportion of cement needed for adequate hardening of the block was also studied. Molasses- and date syrup-based blocks hardened equally well. However, the higher cost of date syrup precluded its use. Wheat straw yielded a low-density block that hardened slowly. Date fibre retained moisture and hardened extremely slowly. Wheat bran-based blocks hardened quickly and yielded dense blocks. Hence, wheat bran was judged to be the superior source of fibre. Lime did not effectively bind the blocks. A cement content of 15% allowed hardening of the blocks within 2-3 weeks. A level of 10% cement in the block reduced the hardening rate by about 50%. Sheep and goats consumed both the straw- and wheat bran-based blocks but at different rates. Consumption of the straw-based block by sheep ranged from 50 to 179 g/head per day, whereas the denser wheat bran-based block was consumed at a rate of 8-20 g/head per day. Consumption of the straw-based block by goats was low (8 g/head per day) compared to that of wheat bran-based blocks (16-24 g/head per day). On the basis of the intake of the bran-based block by sheep, a block was designed that would provide approximately 50% of an animal's trace mineral requirements per day. This block consisted of 45% molasses, 10% urea, 5% trace minerals, 2.5% NaCl, 22.5% wheat bran and 15% cement. Sheep consuming this block gained more weight than sheep fed a conventional mineral block or sheep receiving no mineral supplementation. MUBs are inexpensive (9.5 US cents/kg). We conclude that MUBs have utility for providing trace elements in ruminant diets.

Animal Nutritional Physiological Phenomena↗

beta-Carotene production in sugarcane molasses by a Rhodotorula glutinis mutant.

Several wild strains and mutants of Rhodotorula spp. were screened for growth, carotenoid production and the proportion of -carotene produced in sugarcane molasses. A better producer, Rhodotorula glutinis mutant 32, was optimized for carotenoid production with respect to total reducing sugar (TRS) concentration and pH. In shake flasks, when molasses was used as the sole nutrient medium with 40 g l(-1) TRS, at pH 6, the carotenoid yield was 14 mg l(-1) and -carotene accounted for 70% of the total carotenoids. In a 14-l stirred tank fermenter, a 20% increase in torulene content was observed in plain molasses medium. However, by addition of yeast extract, this effect was reversed and a 31% increase in -carotene content was observed. Dissolved oxygen (DO) stat fed-batch cultivation of mutant 32 in plain molasses medium yielded 71 and 185 mg l(-1) total carotenoids in double- and triple-strength medium, respectively. When supplemented with yeast extract, the yields were 97 and 183 mg l(-1) total carotenoid with a 30% increase in -carotene and a simultaneous 40% decrease in torulene proportion. Higher cell mass was also achieved by double- and triple-strength fed-batch fermentation.

Biomass↗

Simultaneous ethanol and bacterial ice nuclei production from sugar beet molasses by a Zymomonas mobilis CP4 mutant expressing the inaZ gene of Pseudomonas syringae in continuous culture.

AIM: The aim of this work was to construct a Zymomonas mobilis mutant capable of simultaneous ethanol and ice nuclei production from agricultural by-product such as sugar beet molasses, in steady-state continuous culture. METHODS AND RESULTS: A sucrose-hypertolerant mutant of Z. mobilis strain CP4, named suc40, capable of growing on 40% (w/v) sucrose medium was isolated following N-methyl-N'-nitro-N-nitrosoguanidine treatment. Plasmid pDS3154 carrying the inaZ gene of Pseudomonas syringae was conjugally transferred and expressed in suc40. The potential for simultaneous ethanol and bacterial ice nuclei production was assessed in steady-state continuous cultures over a range of dilution rates from 0.04 to 0.13 h(-1). In addition, the fatty acid and phospholipid profile of the three strains was also investigated. Ethanol production up to 43 g l(-1) was achieved at dilution rates below 0.10 h(-1) in sugar beet molasses. Ice nucleation activity gradually increased with increasing dilution rate and the greatest activity, -3.4 log (ice nuclei per cell), was observed at the highest dilution rate (0.13 h(-1)). Both mutant strains displayed a different fatty acid and phospholipid profile compared with the wild-type strain. CONCLUSIONS: The ability of the mutant and recombinant plasmid-containing strains to grow on high sugar concentrations and in high osmotic pressure environments (molasses) can be attributed to their phospholipid and fatty acid contents. SIGNIFICANCE AND IMPACT OF THE STUDY: Taking into account that sugar beet molasses is a low cost agricultural by-product, the simultaneous ethanol and bacterial ice nucleation production achieved under the studied conditions is considered very promising for industrial applications.

Bacterial Outer Membrane Proteins↗

Production of PHB by a Bacillus megaterium strain using sugarcane molasses and corn steep liquor as sole carbon and nitrogen sources.

Poly(hydroxybutyric acid) (PHB) and other biodegradable polyesters are promising candidates for the development of environment-friendly, totally biodegradable plastics. The use of cane molasses and corn steep liquor, two of the cheapest substrates available in Egypt, may help to reduce the cost of producing such biopolyesters. In this work, the effect of different carbon sources was studied. Maximum production of PHB was obtained with cane molasses and glucose as sole carbon sources (40.8, 39.9 per mg cell dry matter, respectively). The best growth was obtained with 3% molasses, while maximum yield of PHB (46.2% per mg cell dry matter) was obtained with 2% molasses. Corn steep liquor was the best nitrogen source for PHB synthesis (32.7 mg per cell dry matter), on the other hand, best growth was observed when ammonium chloride, ammonium sulphate, ammonium oxalate or ammonium phosphate were used as nitrogen sources.

Bacillus megaterium↗

A marked enhancement in phytase production by a thermophilic mould Sporotrichum thermophile using statistical designs in a cost-effective cane molasses medium.

AIMS: Statistical optimization of phytase production by a thermophilic mould Sporotrichum thermophile in a cost-effective cane molasses medium. METHODS AND RESULTS: Sporotrichum thermophile secreted phytase in cane molasses medium at 45 degrees C and 250 rev min(-1) after 5 days. The important factors identified by Plackett-Burman design (magnesium sulfate, Tween 80, ammonium sulfate and incubation period) were further optimized by response surface methodology (RSM). An overall 107% improvement in phytase production was achieved due to optimization. Supplementation of the medium with inorganic phosphate repressed the enzyme synthesis. When inorganic phosphate was reduced from the cane molasses medium by treatment with calcium chloride, the enzyme production increased. The phytase activity was not affected by the enzyme treatment with trypsin and pepsin. CONCLUSIONS: A twofold increase in phytase production was achieved due to optimization using statistical designs in a cost-effective cane molasses medium. SIGNIFICANCE AND IMPACT OF THE STUDY: Phytase production was doubled due to optimization. The enzyme, being resistant to trypsin and pepsin, thermostable and acid stable, can find application in animal feed industry for improving nutritional status of the feed and combating environmental phosphorus pollution.

6-Phytase↗