Two experiments were conducted to investigate the concept that the addition of corn expressing an Escherichia coli-derived gene (corn-based phytase; CBP) to a P-deficient diet would improve growth performance and P utilization in pigs. An E. coli-derived microbial phytase (expressed in Pichia pastoris) sprayed onto a wheat carrier (Quantum) was included for comparison. In Exp. 1, forty-eight 10-kg pigs were blocked by BW into 6 blocks and allotted to 8 dietary treatments such that the BW among dietary treatments was similar and given free access to feed for 28 d. The dietary treatments were a negative control (NC) with no inorganic P supplementation; NC + 2, 4, or 6 g of monosodium phosphate/kg; NC + 16,500, 33,000, or 49,500 phytase units (FTU) of CBP/kg; and NC + 16,500 FTU of Quantum/kg. In Exp. 2, twenty-four 13-kg barrows were assigned to the NC, NC + 16,500 or 33,000 FTU of CBP/kg, or NC + 16,500 FTU of Quantum/kg, in a nutrient- and energy-balance study consisting of 5 d of adjustment and 5-d collection periods. The total collection method was used to determine nutrient and energy balance. Addition of CBP to the low-P NC diet linearly increased (P< 0.01) ADG, G:F, and plasma P concentration of pigs during the 28-d study. There was no difference in ADG, G:F, or plasma P concentration between pigs fed the CBP or Quantum phytase at 16,500 FTU/kg. Weight gain, G:F, and plasma P concentration of pigs increased (P < 0.01) with monosodium phosphate supplementation, confirming P deficiency of the NC diet. Linear improvements (P < 0.05) in DM digestibility and energy retention were observed with CBP supplementation of the NC diet. Although there were linear (P < 0.01) and quadratic (P < 0.05) increases in N digestibility, N retention was unaffected by CBP supplementation of the NC diet in growing pigs. Phosphorus and Ca digestibilities and retentions improved linearly and quadratically (P < 0.01) with the addition of CBP to the NC diet. There was no difference in digestive utilization of P or Ca between pigs fed CBP and Quantum phytase at 16,500 FTU/kg. The data showed that the addition of a corn expressing an E. coli-derived gene to a P-deficient diet improved growth performance and indices of P utilization in pigs, and corn expressing phytase was as efficacious as Quantum phytase when supplemented in P-deficient diets for weanling pigs.

Two experiments were conducted to evaluate the nutritional quality of 2 varieties of Purdue high-digestible sorghum (PHD1 and PHD2) and a normal sorghum, compared with corn, in diets of pigs and broiler chicks. In Exp. 1, 12 pigs (average BW, 55 kg) fitted with ileal T-cannula were fed 4 diets containing 946 g of corn or sorghum (PHD1, PHD2, and normal) per kg in a 2-period crossover design (i.e., each pig received 2 diets over a 2-wk period with 6 pigs per dietary treatment) to determine apparent ileal or total tract digestibility of nutrients and energy. There was no difference in the ileal or total tract digestibility of DM, energy, P, Ca, or N among dietary treatments. In Exp. 2, a total of 192 broiler chicks were grouped by weight into 8 blocks of 4 cages each with 6 chicks per cage, and cages were assigned randomly to 1 of the 4 dietary treatments within each block. Chicks were fed corn-soybean meal (SBM) or sorghum-SBM diets for 21 d to determine apparent total tract retention and then switched to diets containing 935 g of the corresponding corn or 1 of the 3 sorghum varieties per kg for 7 d to determine apparent ileal digestibility and total tract retention. Apparent ileal digestibilities of DM and P, as well as energy, were not different in chicks fed diets

We describe a CCCH type of zinc finger domain in a replication protein A (RPA) homolog found in members of different lineages of the Euryarchaeota, a subdomain of Archaea. The zinc finger is characterized by CX 2 CX 8 CX 2 H, where X is any amino acid. Using MacRPA3, a representative of this new group of RPA in Methanosarcina acetivorans, we made two deletion mutants: a C-terminal deletion mutant lacking the zinc finger and an N-terminal deletion mutant containing the zinc finger domain. Whereas the N-terminal deletion mutant contained zinc at a level comparable to the wild-type protein level, the C-terminal deletion mutant was devoid of zinc. We further created four different mutants of MacRPA3 by replacing each of the four invariable amino acids in the zinc finger with alanine. Each single mutation at an invariable position resulted in a protein containing less than 35% of the zinc found in the wild-type protein. Circular dichroism spectra suggested that although the mutation at the first cysteine resulted in minor perturbation of protein structure, mutations at the other invariable positions led to larger structural changes. All proteins harboring a mutation at one of the invariable positions bound to single-stranded DNA weakly, and this translated into reduced capacity to stimulate DNA synthesis by M. acetivorans DNA polymerase BI. By subjecting the protein and its mutants to oxidizing and reducing conditions, we demonstrated that ssDNA binding by MacRPA3 may be regulated by redox through the zinc finger. Thus, the zinc finger modules in euryarchaeal RPA proteins may serve as a means by which the function of these proteins is regulated in the cell.

The efficacy of corn expressing an Escherichia coli-derived gene (corn-based phytase; CBP) and an E. coli-derived microbial phytase (expressed in Pichia pastoris) sprayed onto a wheat carrier (Quantum) was comparatively evaluated in two 14-d broiler chicken studies. In experiment 1, a total of 288 seven-day-old male broiler chicks were grouped by weight into 8 blocks of 6 cages, with 6 birds per cage, and used to measure growth performance, bone mineralization, and nutrient utilization. In experiment 2, a total of 192 seven-day-old male broiler chicks were used, with 4 birds per cage. Six dietary treatment groups were randomly allotted to the cages within each of 8 blocks. The corn-soybean meal-based diets used in each study consisted of a positive control adequate in P and Ca; a negative control (NC) low in P and Ca (no added inorganic P); the NC supplemented with 3,630, 36,300, or 363,000 phytase units (FTU) of CBP/kg; and the NC + 3,630 FTU of Quantum/kg. Growth performance and bone mineralization criteria were reevaluated in the second broiler study. Data from the 2 experiments were combined because there was no diet x experiment interaction and analyzed as a randomized complete block design. Weight gain decreased (P < 0.05) with a reduction in dietary P and Ca in the NC diet. Weight gain, feed efficiency, and percentage of tibia ash of birds fed 3,630 FTU/kg of either CBP or Quantum phytase were not different. There was a quadratic response (P < 0.01) to CBP supplementation of the NC diets in weight gain of the broiler chicks. Feed intake increased quadratically (P < 0.01) with CBP supplementation of the NC diets, but CBP supplementation of the NC diets did not affect feed efficiency. Tibia ash of birds fed the positive control diet was greater (P < 0.01) than that of birds fed the NC diet. There was a quadratic response (P < 0.01) to CBP supplementation of the NC diets in tibia ash of the broiler chicks. Birds fed supplemental phytase had greater (P < 0.01) ileal and total tract P and Ca digestibility than birds fed the NC diet. Data from the study suggest that E. coli phytase expressed in corn is efficacious in P-deficient broiler chick diets for the improvement of growth performance and indices of P utilization, which would minimize the need for supplemental P in broiler diets.

The residual phytase activity, phytic acid P content, and microstructure of the digesta along the gastrointestinal tract (GIT) of broiler chickens fed a corn expressing microbial phytase was studied in a 14-d experiment. The phytase activity of the corn expressing phytase (CBP) was determined to be 660 phytase units/g and was incorporated into broiler diets at varying rates. One hundred forty-four 7-d-old male broiler chickens were grouped by weight into 8 blocks of 3 cages with 6 birds per cage. Three dietary treatments were randomly allotted to the cages within blocks. The corn-soybean meal-based diets consisting of low P and Ca (no added inorganic P) supplemented with 0 (control), 55, or 550 g/kg of CBP (substituting corn) were formulated to contain 0 (control), 36,300, or 363,000 phytase units/kg of phytase activity, respectively. Birds were fed the dietary treatments for 14 d when they were killed and digesta samples from the proventriculus and gizzard, jejunum, and ileum were collected. The residual phytase activity along the GIT increased linearly and quadratically (P < 0.01) with the addition of CBP to the control diets. There was a decrease (P < 0.01) in residual phytase activity as digesta moved distally along the GIT with CBP supplementation. Phytic acid P content significantly decreased (linear, P < 0.01; quadratic, P < 0.05) with CBP supplementation of the control diets. There was substantial degradation (linear and quadratic, P < 0.01) of phytic acid content caudally along the GIT of birds. Extensive cell wall degradation of digesta samples from the proventriculus and gizzard in broilers fed the highest level of CBP compared with birds fed the control diets was observed. The addition of CBP to control diets led to a rapid degradation of the cell walls of digesta and a marked reduction in phytic acid P concentration of digesta in broiler chicks.