饲料加工工艺对不同剂型植酸酶加工损耗的影响

试验旨在研究热处理对2个进口品牌和2个国产品牌耐高温植酸酶的影响及挤压膨化加工工艺(140~175℃)、环膜硬颗粒加工工艺(70~90℃)、冷挤压平模硬颗粒加工工艺(60~66℃)对不同剂型植酸酶活性损耗率的影响。不同品牌耐高温植酸酶在80℃水浴中处理2 min后测定其活性,同时在饲料中添加4 000、5 000 FTU/kg和8 000 FTU/kg活性的普通酸性植酸酶和耐高温植酸酶,分别测定植酸酶在不同制粒加工工艺条件下的活性。结果表明:热处理对不同耐高温植酸酶活性损耗的影响不同,其活性损耗率范围为8.45%~80.07%。在挤压膨化加工工艺条件下,两种植酸酶基本全部失活;在环模硬颗粒加工工艺条件下,普通商品酸性植酸酶活性损耗率为77.71%,而耐高温植酸酶活性损耗率为34.81%;在冷挤压平模硬颗粒加工工艺条件下,普通商品酸性植酸酶活性损耗率为27.40%,耐高温植酸酶活性损耗率为21.15%。采用内添加方式添加植酸酶,不同加工工艺对植酸酶活性损耗率的影响由大到小依次为:挤压膨化加工工艺环膜硬颗粒加工工艺冷挤压平模硬颗粒加工工艺。     

液体植酸酶的稳定性研究

此文研究了液体植酸酶在不同贮存温度下的酶活稳定性,以及不同稀释倍数和调节稀释后酶液的pH时液体棱酸酶的活性和物理性状的影响,同时探讨了液体植酸酶与液体木聚糖酶复配的稳定性能。试验结果表明：液体植酸酶在低温下贮存酶活稳定性较好;不同稀释倍数对液体植酸酶活性无明显影响,但稀释倍数较大显著影响？植酸酶溶液的物理性状;通过调节稀释后的植酸酶溶液的pH可以显著改善其物理性状。此外,与液体木聚糖酶复配无论是复配后酶液蛇活性还是其物理性状均具有很好盼稳定性。     

植酸酶在肉仔鸡消化道内环境下活性变化的体外研究

本研究在体外条件下对肉仔鸡消化道内pH环境以及蛋白质水解酶对黑曲霉植酸酶活性的影响进行了研究。结果表明,该植酸酶为酸性植酸酶,最适pH为5.5。该植酸酶对pH环境和蛋白质水解酶较为敏感。当环境pH远离其最适pH时其活性迅速下降,且大部分酶活损失发生在30 min内,然后趋于稳定。在pH3.0和pH6.0条件下,加入胃蛋白酶和胰酶后,植酸酶活性亦进一步迅速降低,然后趋于稳定,但与胃蛋白酶相比,胰酶对植酸酶活性的影响更大。从植酸酶在14、28和42日龄肉仔鸡胃和小肠稳定性的研究可以看出,植酸酶在胃存留率分别为42%、40%和49%,在小肠内的存留率仅19%。植酸酶在胃内的稳定性好于小肠,小肠内环境不利于植酸酶存活。植酸酶活性受肉仔鸡消化道pH和蛋白酶的影响较大,这可能是影响植酸酶作用效果的主要因素之一。     

模拟高温制粒过程中酶活损失的快速评定方法

试验采用湿热法,并用水浴和烘箱2种加热方式,模拟在制粒过程中,温度变化对植酸酶酶活损失的变化情况的影响.从市场上收集到A、B、C、D和E5种耐高温植酸酶产品,分别经85℃烘箱处理和85℃水浴处理后,发现烘箱处理组酶A和B活性损失率显著低于水浴处理组,酶D活性损失率显著小于水浴处理组；酶C和E组间活性损失率差异不显著.5种植酸酶分别经85、90和95℃水浴处理后,植酸酶A的活性损失率分别为50.39％、65.64％和71.15％；植酸酶B的活性损失率分别为47.66％、69.90％和73.65％；植酸酶C的活性损失率分别为2.96％和15.46％和29.83％；植酸酶D的活性损失率分别为4.68％、40.45％和47.18％,植酸酶E的活性损失率分别为28.29％、69.11％和71.89％.试验结果表明植酸酶C具有最好的耐热性；采用水浴加热方式的湿热法更适用于试验.     

植物性植酸酶及其应用

本文综述了植物性植酸酶的来源、特征、活性、对植酸磷的分解作用、影响植物性植酸酶活性大小的因素、植物性植酸酶与其他养分之间的关系、从麸皮中提取植酸酶的研究现状等，最后对植物性植酸酶在畜牧业中的应用进行了展望。     

植酸酶应用的影响因素

１饲料加工温度植酸酶水解植酸或植酸盐使其中的磷酸根离子释放出来，其酶的活性随着温度升高而增加。但其作为一种生物活性蛋白质，当温度升至７０℃以上时，酶的活性会因其变性而降低，作为饲料添加剂应用时，人们考虑更多的是饲料加工工艺参数中的制粒温度对植酸酶活性的影响。尽管微生物来源的植酸酶比其他酶制剂更耐高温（植酸酶的最适温度可高达６０～７０℃），但高温度调制过程中的活性损失是在所难免的。解决这地问题的途径有三个，一是特异微生物菌株的筛选，使用Aspergillusniger生产的植酸酶在９０℃环境下放置３０min，其活性…     

瘤胃微生物植酸酶的分类与酶学性质研究进展

反刍动物能很大程度上依靠瘤胃微生物来消化利用饲料中的植酸盐,但目前国内有关瘤胃微生物植酸酶的研究报道较少.本文对瘤胃微生物植酸酶的主要来源和活性进行了综述分析,发现主要有5种瘤胃细菌能分泌植酸酶,其中,反刍兽新月形单胞菌(Selenomonas ruminantium)分泌的植酸酶活性最高(199.1～703.6 U·mL-1)；根据酶促反应的最适pH和催化机理,可将瘤胃微生物植酸酶分别归类为酸性植酸酶和半胱氨酸磷酸酶植酸酶；根据植酸酶水解植酸时的立体专一性,发现瘤胃微生物植酸酶中可能同时存在3-植酸酶和5-植酸酶2类植酸酶.最后本文论述了瘤胃微生物植酸酶酶促反应动力学特性,发现反刍兽新月形单胞菌植酸酶的最适pH为4.0～5.5、最适温度为50～55℃、对植酸和ATP表现出较强的活性；埃氏巨形球菌植酸酶的最适pH为5,0、最适温度为60℃、仅对植酸表现出活性.     

无淀粉麦麸为底物体外植酸酶活性评价方法的建立

为提高体外检测方法的科学性和客观性,预测植酸酶在动物体内的活性和作用效果,研究以无淀粉麦麸为底物,探索并建立了一种更为客观评价植酸酶活性的检测方法.与现有的植酸酶酶活测定国家标准相比,在不同pH值的缓冲液中检验植酸酶降解天然植酸的效果,具有更为科学、客观与简便等优点.     

植物性植酸酶及其应用

许多作物籽实及其加工副产品中均含有天然植酸酶,如小麦、玉米、大麦、黑麦、小黑麦、燕麦、水稻、豆类、麸皮等,籽实中的植酸酶已被分离鉴定。不同作物、不同品种植酸酶的含量(活性)差异很大,玉米、高粱、油菜籽中的植酸酶活性很低,而麦类及一些蒸馏副产品(如玉米蒸馏物)均含有很高的植酸酶活性。     

新型耐热植酸酶对肉仔鸡生长性能及血清生理生化指标的影响

旨在通过饲喂肉仔鸡添加普通植酸酶与耐高温植酸酶的日粮,比较2种植酸酶对肉仔鸡生产性能及血清生理生化指标的影响。选用4 800只1日龄AA肉仔鸡,随机分为6个处理,每处理5个重复。试验分2期进行,1～3和4～6周。试验对照组饲喂基础日粮,试验1组降低基础日粮磷的水平,试验2组与3组分别在试验1组的基础上添加普通植酸酶500和2 000U.kg-1,试验4组和5组分别添加耐高温植酸酶250和500U.kg-1。结果表明:(1)当降低基础日粮中磷添加水平时,日增重显著下降(P<0.05),料重比提高;在此基础上添加植酸酶,日增重提高,料重比下降,甚至优于对照组。从生长性能指标分析来看,250～500U.kg-1耐热植酸酶与2 000U.kg-1普通植酸酶效果相当,优于试验2组;(2)当降低基础日粮中磷添加水平时,血清磷水平显著降低,血清碱性磷酸酶活性显著提高;在此基础上添加植酸酶,血清磷水平和碱性磷酸酶活性恢复到对照组水平。对血清总蛋白、球蛋白、白蛋白、谷草转氨酶、谷丙转氨酶和钙含量等指标含量基本没有影响;(3)不同日粮处理对T4、T3、生长激素、胰岛素样生长因子Ⅰ、甲状旁腺激素和降钙素均没有显著性影响(P>0.05);与对照组相比,胰岛素样生长因子Ⅰ在试验组1呈降低趋势(P>0.05),而各植酸酶添加组呈提高趋势。本试验条件下,发现在低磷日粮中添加新型耐热植酸酶可以显著提高肉仔鸡的平均日增重,降低料重比;对于血清生理生化指标,2种植酸酶添加效果无显著差异。     

Comparative enzymatic hydrolysis of phytate in various animal feedstuff with two different phytases

Bacillus amyloliquefaciens DS11 phytase (DS11 phytase) and Aspergillus ficuum phytase (AF phytase) activities were investigated by measuring the release of phosphate from phytate in animal feedstuff such as wheat bran, corn meal, soybean meal and rice flour at pH 5 and 7. In all the tested feedstuff, the enzymatic activity of DS11 phytase was more active at pH 7, but that of AF phytase was more active at pH 5. From these results, the phytate in the gastrointestinal tract could be degraded in the small intestine or stomach by DS11 or AF phytase, respectively. In conclusion, the results presented in this paper indicated that different combination ratios of DS11 and AF phytase, depending on the kind of feedstuff, might effectively induce more enzymatic activity both in the stomach and small intestine in terms of the pH of the gastrointestinal tract.     

Factors affecting storage stability of various commercial phytase sources

A 360-d study was performed to evaluate the effects of different environmental conditions on storage stability of exogenous phytases. Coated and uncoated products from 3 phytase sources [Ronozyme P (DSM Nutritional Products, Basel, Switzerland), OptiPhos (Phytex LLC, Sheridan, IN), and Phyzyme (Danisco Animal Nutrition, Marlborough, UK)] were stored as pure forms, in a vitamin premix, or in a vitamin and trace mineral (VTM) premix. Pure products were stored at -18, 5, 23, and 37°C (75% humidity). Premixes were stored at 23 and 37°C. Sampling was performed on d 0, 30, 60, 90, 120, 180, 270, and 360. Sampling of the pure products stored at -18 (lack of sample) and 5°C (because of mold growth) was discontinued after d 120. Stability was reported as the residual phytase activity (% of initial) at each sampling point. For the stability of the pure forms, all interactive and main effects of the phytase product, coating, time, and storage temperature were significant (P < 0.01), except for the time × coating interaction. When stored at 23°C or less, pure phytases retained at least 91, 85, 78, and 71% of their initial phytase activity at 30, 60, 90, and 120 d of storage, respectively. However, storing pure products at 37°C reduced (P < 0.01) phytase stability, with OptiPhos retaining the most (P < 0.01) activity. Coating mitigated (P < 0.01) the negative effects of high storage temperature for Ronozyme and OptiPhos (from d 90 onward), but not for Phyzyme. For the stability of phytase in different forms of storage, all interactive and main effects of phytase product, form, coating, time, and temperature of storage were significant (P < 0.01). When stored at room temperature (23°C), retained phytase activities for most the phytase sources were more than 85, 73, and 60% of the initial activity up to 180 d when stored as pure products, vitamin premixes, or VTM premixes, respectively. When stored at 37°C, pure phytase products had greater (P < 0.01) retention of initial phytase activity than when phytases were mixed with the vitamin or VTM premixes. Coated phytases stored in any form had greater (P < 0.01) activity retention than the uncoated phytases at all sampling periods. Results indicate that storage stability of commercially available phytases is affected by duration of storage, temperature, product form, coating, and phytase source. Pure products held at 23°C or less were the most stable. In premixes, longer storage times and higher temperatures reduced phytase activity, but coating mitigated some of these negative effects.     

Improvement of growth and nutritive value in chicks with non-genetically modified phytase product from Aspergillus niger

1. Non-genetically modified (non-GM) phytase product derived from Aspergillus niger possesses various side active enzymes including α -amylase, protease, cellulase and hemicellulase. In contrast, the product of genetically modified (GM) phytase product has much less side active enzyme since the capacity of phytase production is reinforced by gene modification. In the present study we have tried to determine whether the difference of side enzyme activity of phytase product affects growth performances and nutritive value in chicks; in addition we tried to characterise the physiological change induced by the difference of side active enzymes. 2. Single Comb White Leghorn male chicks at 7 d of age were fed on experimental barley-based diets for 10 d. The feeding trial was of a factorial design (3 × 2 × 2), having three types of dietary phytase products (control, non-GM or GM phytase products derived from A. niger at 1000 U/kg diet), two levels of dietary available P supplement (0 or 6 g/kg diet) and two levels of dietary protein (CP 180 or 120 g/kg). 3. The non-GM phytase product caused a 6% increase in final body weight and feed efficiency com6 pared with the control and the GM phytase product without interacting with dietary protein and available P level. However, in birds given available P-free diet, both non-GM and GM phytase products induced a 20% increase in plasma P concentration, suggesting no difference in phytase activity between the non-GM and GM phytase products. 4. The balance study showed that the metabolisable energy of the non-GM phytase product (15.6 ± 0.05 kJ/g diet) was significantly higher among the treatments (control, 15.1 ± 0.05; GM phytase product 15.3 ± 0.07). The non-GM phytase product also increased the rate of food passage through the crop, and caused a drastic reduction in intestinal weight, perhaps as a consequence of digestion of non-starch polysaccharides. 5. We conclude that the side active enzymes in non-GM phytase product improve growth performance and nutritive value of the diet in chicks. However, the efficacy of phytase activity should not be different between non-GM and GM phytase products.     

Comparison of Aspergillus niger phytase and Trichoderma reesei phytase and acid phosphatase on phytate phosphorus availability in pigs fed on maize-soybean meal or barley-soybean meal diets.

The efficacy of Aspergillus niger (APhy) phytase, Trichoderma reesei (TPhy) phytase and acid phosphatase (TAcPh) preparations in improving the utilization of phytin-phosphorus in the maize-soybean meal (SBM) or barley-SBM (800:200 g kg-1) diets was studied in two separate digestibility and balance trials with ten growing pigs using 5 x 5 Latin square designs. The positive control diet contained a total phosphorus (P) of 6.5 g kg-1, while the negative control as well as the APhy, TPhy and TAcPh supplemented diets which did not contain additional inorganic-P, had a total P of 4.1 g kg-1. The APhy and TPhy supplements provided phytase activity of 1000 PU g-1 together with AcPh of 8000 HFU g-1. TAcPh at a level of 8000 HFU g-1 was the only addition to one diet. The intrinsic phytase activity of barley was 355 PU g-1 while maize and soybean meal showed no phytase activity. Phytase supplements of the APhy and TPhy sources increased ash digestibility in both diets but had only a minor effect on nitrogen utilization. The addition of phytase improved absorption of P by 21%-units in barley-SBM diet and 29%-units in maize-SBM diet, without any difference between the two phytase sources. The retained P in diets with phytase was higher than in diets without phytase, 4.4 (APhy), 4.5 (TPhy) vs. 2.9 g d-1 in barley-SBM-diets and 3.7 (APhy), 4.0 (TPhy) vs. 1.8 g d-1 in maize-SBM-diets. No difference was found between the two sources of phytase. TAcPh without additional phytase did not show any effect on P absorption or retention. Ca absorption and retention were improved due to the phytase treatments. Supplementing pig diets with either APhy or TPhy sources seems to be equally effective in enhancing the availability of phytate-P. Consequently, these supplements can reduce the P-excretion of pigs by 32-40% as compared with the diet supplemented with inorganic-P.     

Effect of diet containing phytate and phytase on the activity and messenger ribonucleic acid expression of carbohydrase and transporter in chickens

The effect of dietary phytate and phytase on carbohydrase activity and hexose transport was investigated in broiler chickens. Diets containing phytate P (2.2 or 4.4 g/kg) with different phytase dose rates (0, 500, or 1,000 phytase units/kg) were fed to 504 female Cobb chicks for 3 wk. Diets containing high phytate concentrations depressed (P < 0.05) BW and G:F, whereas phytase supplementation improved (P < 0.05) the performance of birds. In the duodenum, phytate decreased (P < 0.05) the activities of disaccharidases, Na(+)K(+)-ATPase, and glucose concentrations by 5 to 11%, but phytase enhanced (P < 0.05) the concentrations of amylase, sucrase, maltase, Na(+)K(+)-ATPase, and glucose by 5 to 30%. In the jejunum, phytate decreased (P < 0.05) the concentrations of amylase, sucrase, Na(+)K(+)-ATPase, and glucose by 10 to 22%, and phytase alleviated the negative effect of phytate on the above variables. Ingestion of diets containing phytate also decreased (P < 0.05) serum amylase activity and glucose concentration, and phytase enhanced (P < 0.05) serum concentrations of amylase, sucrase, maltase, Na(+)K(+)-ATPase, and glucose. There were also interactions (P < 0.05) between phytate and phytase on the concentrations of serum amylase, duodenal amylase, sucrase, and jejunal glucose. Enzymatic analysis at a molecular level showed that neither phytate nor phytase influenced the mRNA expression of sucrase-isomaltase in the small intestine. Also, the investigation into the sodium glucose cotransporter gene may challenge the mechanism by which phytate interferes with glucose utilization, as partly indicated by bird performance, and transmembrane transport because diets containing increased phytate upregulated (P < 0.05) the mRNA expression of the sodium glucose cotransporter gene in duodenum and did not influence it in the jejunum. These results indicate that phytate can impair endogenous carbohydrase activity and digestive competence, and phytase can ameliorate these effects for chickens.     

Evaluation of phytase concentration needed for growing-finishing commercial turkey toms

1. Growth performance, serum bone markers, and bone strength and mineralisation were determined in tom turkeys grown from 9 to 17 weeks of age. 2. Dietary non-phytate phosphorus was formulated to be reduced by 1.0 g/kg in the low phosphorus diet compared to a control diet and phytase was added to provide 0, 150, 300, 450 or 600 units/kg activity to the low phosphorus diet. 3. From 9 to 12 weeks of age, body weight and gain:food were reduced by the low phosphorus diet without added phytase, compared to the adequate phosphorus diet. Increasing the concentration of phytase linearly increased these growth parameters. There were no significant growth responses at 17 weeks of age. 4. Serum osteocalcin was reduced by increasing dietary phosphorus at 12 weeks of age when growth was affected, but not at later ages. Serum pyridinoline was reduced by higher dietary phosphorus and decreased linearly with increasing phytase activity at 17 weeks of age. 5. Fracture force of the ulna and femur increased linearly with increasing phytase activity but bone strength was not affected when corrected for bone cross-sectional area. Bone strength of the ulna and ash concentration of the ulna and tibia were increased by higher dietary phosphorus. Humerus and ulna ash increased linearly with increasing phytase activity. 6. Water-soluble phosphorus content of the litter was increased by higher dietary phosphorus and addition of phytase to the low phosphorus diet. The increase in water-soluble phosphorus content of the litter when phytase was fed may indicate that phosphorus could be fed at a lower concentration than used in this trial, at least in the finisher diet when phytase is added to the food. 7. Bone fracture force, strength and ash were generally optimised when 450 units/kg phytase activity was added to the low phosphorus diet. However, growth performance was best in the grower I (9 to 12 weeks) phase when 600 units/kg phytase was added to the diet.     

An improved method for a rapid determination of phytase activity in animal feed

The current direct colorimetric assay for phytase activity in feeds has interference from high P background and other factors. Our objective was to develop a rapid and reliable spin column method to accurately determine phytase activity in feed ingredients or complete diets. After the feed sample was extracted by stirring in 0.2 M citrate buffer, pH 5.5, for 30 min at room temperature, the oily layer of the supernatant fraction was removed by passing through an acrodisc syringe filter (0.45-microm HT Tuffryn membrane, Gelman Laboratory, Ann Arbor, MI). The filtrate was then loaded onto a spin column (MW cutoff 30,000, Millipore, Bedford, MA) to remove free phosphate before the phytase activity assay. Compared with the direct assay, this new procedure improved both accuracy and reproducibility. When diets contained phytase at 0 to 1,500 U/kg (as fed), the CV for multiple assays of the same samples (n = 6) by the new method ranged from 1 to 6% compared with 28 to 39% by the direct method. A linear relationship was found between the added phytase activity in practical diets and the analyzed activity by the new method (r2 = 0.99; P < 0.01). In conclusion, the spin column method is an improved assay for phytase activity in animal feed, and may be used for quality control of phytase supplementation.     

国标法测定植酸酶活性存在问题的探讨

利用国标法测定植酸酶活性过程中发现,植酸钠底物溶液pH为6.48,高于酶活性单位定义中pH5.5。为进一步探讨植酸钠底物溶液pH对植酸酶活性的影响,本试验研究了调节与不调节植酸钠底物溶液pH对样品空白的吸光值以及植酸酶活性的影响。结果表明:将植酸钠溶液pH由6.48调至5.5对酶活测定中的样品空白无显著影响(P>0.05),但极显著影响植酸酶活性(P<0.01),未调节pH组植酸酶活性仅相当于调节pH组的66%,利用不同的酸调节植酸钠溶液pH,对植酸酶活性无显著影响(P>0.05)。     

The effect of previous exposure to dietary microbial phytase on the endogenous excretions of energy, nitrogen and minerals from turkeys

1. A precision feeding experiment was conducted with turkeys, which had previously been fed diets with or without phytase, to study the effects on the excretions of endogenous energy (EEL), nitrogen (ENL), amino acids (EAAL) and minerals. 2. Female turkeys (BUT 6) which had been fed one of 4 experimental diets (low P maize-soya diets (control, C), C + 250 international units of phytase/kg diet (FTU), C + 500 FTU and C + 2500 FTU) were used in this study. All birds were fasted and then given 50 mL of glucose solution at 46 d of age. Birds were allocated to individual metabolism cages in a randomised block design with 8 replicates for each of the 4 previously-fed diets. 3. The response of EEL and ENL to phytase pre-exposure was linear. An increase of 100 FTU reduced the EEL and ENL by approximately 1·6 kJ and 20 mg respectively. The results suggest that a minimum activity of phytase of 500 FTU is needed to initiate the reduction of these losses. 4. Pre-exposure to phytase reduced the EAAL, which was best described as a linear response with increasing phytase dose in the pre-study period. An increase of 100 FTU reduced the losses of total endogenous amino acids by approximately 225 mg. 5. In contrast to the results for endogenous energy losses, turkeys pre-exposed to phytase linearly increased their excretions of Ca and Mg with increasing phytase activity in the pre-study period. 6. The effects of feeding turkeys with supplementary phytase continued for at least 4 d after the diets were withdrawn. This suggests that exposure to phytase alters the functionality and secretions of the gastrointestinal tract, which may influence the nutritive value of diets fed immediately after.     

不同植酸酶对生长育肥猪生产性能及养分利用的影响

选择"杜×长×大"三元杂交生长猪54头,研究了大肠杆菌、黑曲霉来源植酸酶替代日粮中部分磷酸氢钙对生长育肥猪生长性能、养分消化率、血液生化指标及粪磷、氮排泄的影响。试验猪随机分成对照组、试验Ⅰ组、试验Ⅱ组,每组3个重复。对照组饲喂常规日粮,试验Ⅰ组饲喂低磷+植酸酶Ⅰ的日粮,试验Ⅱ组饲喂低磷+植酸酶Ⅱ的日粮。试验结果表明,在生长育肥猪日粮中添加植酸酶替代部分磷酸氢钙,改善了生长育肥猪生长性能,其中添加大肠杆菌植酸酶显著改善生长育肥猪的生长性能;显著改善了干物质、粗蛋白质、钙、磷、总能的表观消化率,并显著降低了粪中氮、磷的排泄;显著提高了血清中总蛋白、球蛋白、血清磷的浓度,而尿素氮、血清钙浓度显著降低。添加黑曲霉植酸酶后,血糖浓度下降,而碱性磷酸酶的活性无显著变化;添加大肠杆菌植酸酶后,碱性磷酸酶的活性显著降低,而血糖浓度无显著变化。