Phytate-degrading enzymes in pig nutrition

Phytate, the mixed salt of phytic acid (myo-inositol hexaphosphate), derived from plant-sourced feed ingredients is invariably present in practical diets for pigs. Typically, swine diets contain in the order of 3.0 g kg^− 1 phytate-bound phosphorus (phytate-P) but phytate concentrations are subject to variation. Importantly, phytate-P is only partially utilised by pigs because they do not generate sufficient endogenous phytase activity. Phytate-degrading enzymes, via step-wise dephosphorylation of phytate, have the capacity to liberate phytate-P, thus enhancing P absorption and reducing P excretion, which are both nutritionally and ecologically beneficial consequences. The commercial introduction of microbial phytases in 1991 has greatly magnified the interest in the roles of phytate and phytase in pig nutrition.

The capacity of microbial phytases to enhance growth performance of pigs offered diets with inadequate P levels is well documented. However, in some instances, phytase has been shown to improve performance of pigs offered P adequate diets thus phytase-induced improvements in growth performance should not be attributed entirely to increased P availability. This raises the possibility that phytase is increasing the utilisation of nutrients other than P. These so-called ‘extra-phosphoric’ effects of phytase remain controversial, particularly in relation to protein and amino acid availability. There are conflicting opinions that are reflected in the inconsistent outcomes of studies to determine the effect of phytase on ileal digestibility of amino acids and protein utilisation in pigs. In phytase amino acid digestibility assays, it seems likely the choice of chromic oxide as the dietary marker has contributed to these ambiguous results, which may be further complicated when ileal digesta samples are taken from cannulated pigs fed on a restricted, twice-daily basis. In order to resolve this critical issue, there is an urgent need to assess the impacts of selection of dietary markers, methods of ileal digesta collection and feeding regimen relative on the outcomes of phytase amino acid digestibility assays in pigs.

However, inconsistent results from phytase studies in pigs are not confined to amino acid digestibility assays. Arguably, insufficient attention has been paid to dietary substrate levels in relation to phytase inclusion from both scientific and practical standpoints. Phytate analyses are not straightforward and there is a real need to develop more accurate and rapid methods to facilitate phytate determinations. The properties of phytate vary between (and within) feed ingredients where solubility of phytate may be critical; which, in turn, is a function of gut pH in pigs. Contemporary phytases have the capacity to degrade approximately 50% of dietary phytate at the level of the ileum, which may mean higher inclusion rates are warranted. Consequently, there is scope for the development of more effective ‘second-generation’ phytate-degrading feed enzymes and their possible introduction, coupled with a better scientific understanding of relevant fundamental issues, will ensure that phytate-degrading enzymes will contribute to viable and sustainable pig production to an even greater extent in the future.     

植酸酶的研究进展

植酸酶是能降解饲料中植酸及其盐的酶。它能提高磷利用率,解除植酸对一些矿物元素如钙、锌、铁、铜等的抗营养效应,不仅对动物具有良好的增重效果,同时可降低动物排泄磷量,有利于环境保护。植酸酶作为饲料添加剂其作用效果受到饲料中钙、磷水平和钙磷比例以及维生素Ｄ含量的影响。植酸酶的运用需要降低饲料中钙、磷水平,维生素Ｄ与植酸酶之间可能存在协同效应。对植酸酶运用的经济分析表明,使用植酸酶能代替饲料中需添加的无机磷。     

Phytate and phytase in fish nutrition

Phytate formed during maturation of plant seeds and grains is a common constituent of plant-derived fish feed. Phytate-bound phosphorus (P) is not available to gastric or agastric fish. A major concern about the presence of phytate in the aquafeed is its negative effect on growth performance, nutrient and energy utilization, and mineral uptake. Bound phytate-P, can be effectively converted to available-P by phytase. During the last decade, phytase has been used by aqua feed industries to enhance the growth performance, nutrient utilization and bioavailability of macro and micro minerals in fish and also to reduce the P pollution into the aquatic environment. Phytase activity is highly dependent on the pH of the fish gut. Unlike mammals, fish are either gastric or agastric, and hence, the action of dietary phytase varies from species to species. In comparison to poultry and swine production, the use of phytase in fish feed is still in an unproven stage. This review discusses effects of phytate on fish, dephytinisation processes, phytase and pathway for phytate degradation, phytase production systems, mode of phytase application, bioefficacy of phytase, effects of phytase on growth performance, nutrient utilization and aquatic environment pollution, and optimum dosage of phytase in fish diets.     

Evaluation of Phytase Enzyme with Chicks Fed Basal Diets Containing Different Soybean Meal Samples

Soybean meal contains approximately 0.62% total P of which 0.4% can be phytate P, which is considered less biologically available for poultry than other forms of P. Soybean meal is a key ingredient in poultry feeds and information is needed about the range of phytate P and nonphytate P in different soybean meals. The phytate P content of soybeans may vary due to climatic conditions, soil type and soybean variety. Previous research has shown that phytate P can be hydrolyzed in the gastrointestinal tract providing available P by adding a commercial phytase enzyme to poultry feed. The extent of phytate hydrolysis by dietary supplementation of phytase has been shown to vary depending on the type of dietary ingredients such as corn, soybean meal, canola meal, and wheat. Research is needed to determine if different commercially available soybean meals respond in a similar manner to a feed added phytase. Twenty-five soybean meal samples were collected from active soybean crushing plants in the United States and 18 of the samples were selected to evaluate the effect of a microbial phytase on phytate P disappearance using 5-d bioassays. The range of analyzed values in soybean meal samples for total P, phytate P, Ca, protein, and neutral detergent fiber (NDF) were 0.59 to 0.87, 0.32 to 0.42, 0.28 to 0.54, 40.44 to 51.69, and 7.78 to 16.09%, respectively. Bioassay results indicate that body weight, feed consumption, and feed conversion ratio improved significantly (P < 0.05) in some of the groups fed diets with enzyme compared with groups fed the same diet with no added enzyme. The range of total P retention and phytate P disappearance for groups fed diets with no enzyme were 21.35 to 48.41 and 13.64 to 37.13%, respectively. The addition of phytase increased total P retention and phytate P disappearance from 56.81 to 68.62 and 76.18 to 94.08%, respectively. The results indicate no correlation among components (total P, phytate P, Ca, protein, and NDF) of soybean meal samples, percentage of phytate P disappearance, and percentage of total P retention for groups fed diets with and without added phytase.     

饲用植酸酶的研究进展

植酸是畜禽植物性饲料中磷的主要存在形式,也是一种广谱性的抗营养因子。植酸酶是催化植酸及植酸盐水解成肌醇与磷酸(盐)的一类酶的总称,可改善磷酸盐的利用率,减轻动物排泄物中的磷对环境的污染,提高营养成分的吸收利用。目前植酸酶在实际应用中还存在着多方面的问题,如表达量低、热稳定性不能满足饲料制粒的要求、作用条件与畜禽体内pH值环境存在差异、对消化酶的抗性差等。基因工程等生物学上游技术的发展,为这些问题的解决提供了有效途径。文章综述了植酸酶基因工程研究的最新进展,并讨论其进一步的研究发展方向。     

Degradation of phytate in the gut of pigs--pathway of gastro-intestinal inositol phosphate hydrolysis and enzymes involved.

The present study gives an overview on the whole mechanism of phytate degradation in the gut and the enzymes involved. Based on the similarity of the human and pigs gut, the study was carried out in pigs as model for humans. To differentiate between intrinsic feed phytases and endogenous phytases hydrolysing phytate in the gut, two diets, one high (control diet) and the other one very low in intrinsic feed phytases (phytase inactivated diet) were applied. In the chyme of stomach, small intestine and colon inositol phosphate isomers and activities of phytases and alkaline phosphatases were determined. In parallel total tract phytate degradation and apparent phosphorus digestibility were assessed. In the stomach chyme of pigs fed the control diet, comparable high phytase activity and strong phytate degradation were observed. The predominant phytate hydrolysis products were inositol phosphates, typically formed by plant phytases. For the phytase inactivated diet, comparable very low phytase activity and almost no phytate degradation in the stomach were determined. In the small intestine and colon, high activity of alkaline phosphatases and low activity of phytases were observed, irrespective of the diet fed. In the colon, stronger phytate degradation for the phytase inactivated diet than for the control diet was detected. Phytate degradation throughout the whole gut was nearly complete and very similar for both diets while the apparent availability of total phosphorus was significantly higher for the pigs fed the control diet than the phytase inactivated diet. The pathway of inositol phosphate hydrolysis in the gut has been elucidated.     

Phytate in pig and poultry nutrition

Phosphorus (P) is primarily stored in the form of phytates in plant seeds, thus being poorly available for monogastric livestock, such as pigs and poultry. As phytate is a polyanionic molecule, it has the capacity to chelate positively charged cations, especially calcium, iron and zinc. Furthermore, it probably compromises the utilization of other dietary nutrients, including protein, starch and lipids. Reduced efficiency of utilization implies both higher levels of supplementation and increased discharge of the undigested nutrients to the environment. The enzyme phytase catalyses the stepwise hydrolysis of phytate. In respect to livestock nutrition, there are four possible sources of this enzyme available for the animals: endogenous mucosal phytase, gut microfloral phytase, plant phytase and exogenous microbial phytase. As the endogenous mucosal phytase in monogastric organisms appears incapable of hydrolysing sufficient amounts of phytate‐bound P, supplementation of exogenous microbial phytase in diets is a common method to increase mineral and nutrient absorption. Plant phytase activity varies greatly among species of plants, resulting in differing gastrointestinal phytate hydrolysis in monogastric animals. Besides the supplementation of microbial phytase, processing techniques are alternative approaches to reduce phytate contents. Thus, techniques such as germination, soaking and fermentation enable activation of naturally occurring plant phytase among others. However, further research is needed to tap the potential of these technologies. The main focus herein is to review the available literature on the role of phytate in pig and poultry nutrition, its degradation throughout the gut and opportunities to enhance the utilization of P as well as other minerals and nutrients which might be complexed by phytates.     

植酸酶在饲料中的应用

植酸酶是一种畜禽饲料添加剂,能有效地降解植酸盐,将植酸磷(六磷酸肌醇)降解为肌醇和无机磷,解除植酸的抗营养作用。植酸酶应用于饲料中,不仅可以提高植物性饲料中植酸磷的消化率,减少无机磷的添加量和磷排泄污染,还可以改善和提高畜禽对饲料中蛋白质、能量、氨基酸和微量元素的利用率,提高动物的生产性能。     

植酸酶在畜禽养殖上的研究进展

植酸酶作为一种畜禽饲料添加剂,能有效地将植酸磷降解为肌醇和无机磷,消除植酸的抗营养作用。本文综述了植酸酶的来源、影响酶作用的因素及其在畜禽生产中的应用。     

饲用植酸酶来源及功能研究进展

植酸酶是一种畜禽饲料添加剂,能有效地将植酸磷降解为肌醇和无机磷,解除植酸的抗营养作用。本文综述了植酸酶的特性、种类、来源及其在动物生产中的应用。     

植酸酶在饲粮中超量添加的研究进展

植酸酶作为常规饲料添加剂已经普遍应用于饲粮中,用于分解饲料原料中的植酸,释放磷元素,降低用于提供有效磷的磷酸氢钙或其他原料的使用量。随着研究的深入,发现植酸酶的添加量还远远达不到其应有的功效,超量添加植酸酶可以分解超过90%的植酸,还可以提高蛋白质、脂肪、淀粉及矿物质元素的利用率,提高畜禽的生产性能,降低饲料成本,减少环境污染。     

Variation in levels of non-starch polysaccharides and endogenous endo-1,4-β-xylanases affects the nutritive value of wheat for poultry

1. Endo-1,4-β-xylanase is known to improve the nutritive value of wheat-based diets for poultry by degrading dietary arabinoxylans. However, broilers’ response to supplementation of wheat-based diets with exogenous endo-1,4-β-xylanase is not always observed.

2. In this study, 108 different wheat lots were analysed for levels of extract viscosity as well as for endogenous endo-1,4-β-xylanase activity, and the impact of these two variables in animal performance was tested.

3. Results revealed that endogenous endo-1,4-β-xylanase activity and extract viscosity content varied widely among different wheat lots. Thus, a trial was conducted to evaluate the efficacy of exogenous enzyme supplementation in broiler diets using wheats with different levels of extract viscosity and endogenous endo-1,4-β-xylanase activity.

4. The data revealed that exogenous enzyme supplementation was only effective when the wheat present in the diet had high levels of extract viscosity (14.8 cP) with low endogenous endo-1,4-β-xylanase activity (347.0 U/kg). Nevertheless, it is apparent that exogenous microbial xylanases reduce digesta extract viscosity and feed conversion ratio independently of the endogenous properties presented by different wheat lots.

5. The data suggest that extract viscosity and/or endogenous endo-1,4-β-xylanase activity affect the response to enzyme supplementation by poultry fed on wheat-based diets.     

Consequences of calcium interactions with phytate and phytase for poultry and pigs

Despite increasing practical experience and cascades of scientific reports on exogenous microbial phytases, several issues associated with their use remain unresolved because of the ambiguous and, at times, conflicting data that has been generated. One possible cause of these inconsistent outcomes is dietary calcium (Ca) levels, which are mainly derived from limestone. Thus the purpose of this review is to examine Ca interactions with dietary phytate and phytases, particularly exogenous, microbial phytases, and their consequences for poultry and pigs. The polyanionic phytate molecule has a tremendous capacity to chelate cations and form insoluble Ca–phytate complexes, which are refractory to phytase activity. Thus Ca–phytate complex formation along the gastrointestinal tract, where one phytate (IP₆) molecule binds up to five Ca atoms, assumes importance and approximately one third of dietary Ca may be bound to phytate in digesta. Consequently, phytate limits the availability of both P and Ca as a result of insoluble Ca–phytate complex formation, the extent of which is driven by gut pH and molar ratios of the two components. It is accepted that Ca–phytate complexes are mainly formed in the small intestine where they have a substantial negative influence on the efficacy of mucosal phytase. However, exogenous phytases are mainly active in more proximal segments of the gut and lower pH levels, so their efficacy should not be influenced by Ca–phytate complexes in the small intestine. There is, however, data to indicate that Ca and phytate interactions occur under acidic conditions with the formation of soluble and insoluble Ca–phytate species, which could negatively impact on exogenous phytase efficacy. Also, Ca will tend to elevate gut pH because of limestone's very high acid binding capacity, which will favour Ca–phytate interactions and may influence the activity of exogenous phytases depending on their pH activity spectrum. The de novo formation of binary protein–phytate complexes that are refractory to pepsin hydrolysis may be fundamental to the negative impact of phytate on the digestibility of protein/amino acids. However, high dietary Ca levels may disrupt protein–phytate complex formation by interacting with both phytate and protein even at acidic pH levels, thereby influencing the outcomes of phytase amino acid digestibility assays. Finally, it is increasingly necessary to define the Ca and nonphytate-P requirements of pigs and poultry offered phytase-supplemented diets.     

影响高粱饲用价值主要内在因素及其对策

大量研究结果显示,饲用高粱含有3种物质不利于高粱在动物体内的消化,一是酚类化合物,二是醇溶蛋白,三是植酸。本文从3类化合物的特征、对消化酶的抑制作用、醇溶蛋白与淀粉以及氨基酸的消化率、植酸和植酸磷以及酶制剂在高粱型畜禽饲粮中的应用等方面,就高粱在动物饲料应用的研究动态和进展进行了简要综述。     

植酸酶及其在畜禽生产中的最新应用研究

植酸酶是一种新型饲料添加剂,它能提高动物对饲料中植酸磷的利用率,降低粪便中磷的排泄量。本文对植酸的化学结构、理化性质与植酸酶的来源、生物学特性,并对影响植酸酶活力的因素及其在畜禽生产中的最新应用研究作了简要介绍。     

Influence of phytate level on broiler performance and the efficacy of 2 microbial phytases from 0 to 21 days of age

Phytate is an antinutrient in animal feeds, reducing the availability and increasing the excretion of nutrients. Phytases are widely used to mitigate the negative influences of phytate. This trial was designed to compare the efficacy of 2 Escherichia coli-derived phytases on broiler performance and bone ash as influenced by dietary phytate level. A total of 1,024 Arbor Acres male broilers were used with 8 replicate pens of 16 birds/pen. Experimental diets were based on low available phosphorus (avP; 1.8 g/kg) with low (6.40 g/kg) or high (10.65 g/kg) phytate. The low-avP diets were then supplemented with mono-dicalcium phosphate to increase the avP level to 4.5 g/kg, 500 phytase units/kg of phytase A, or 500 phytase units/kg of phytase B to create 8 experimental diets. Feed intake, BW gain, FCR, and livability were influenced by a P source × phytase interaction. Feed intake, BW gain, and livability were reduced and FCR was higher in broilers fed low-avP diets, particularly in the presence of high phytate. Phytase A or phytase B improved feed intake, BW gain, and FCR, particularly in the high-phytate diet. However, broilers fed phytase A ate more and were heavier than broilers fed phytase B. Tibia ash was lowest in broilers fed the low-avP diet and highest in broilers fed the diet supplemented with mono-dicalcium phosphate. Phytase increased tibia ash, and broilers fed phytase A had an increase in tibia ash compared with broilers fed phytase B. In conclusion, high dietary phytate reduced broiler performance. Phytase A and phytase B improved bone ash and growth performance, especially in the high-phytate diets. However, phytase A was more efficacious than phytase B, regardless of the level of phytate.     

Relationship between mineral availabilities and dietary phytate in animals

A large amount of phosphorus (P) in corn and soybean meal is in the form of phytate that is poorly available to monogastric animals. It leads to the presence of large amounts of P in manure, which contributes to the P pollution problem. The fermentation of soybean meal with Aspergillus usamii almost completely degraded phytate and improved P availability in chicks. Although dietary yeast phytase increased P absorption and availability in pigs, its efficacy was less than that of Aspergillus niger phytase. It was suggested that the lesser efficacy of yeast phytase resulted from its lower stability against pepsin. Phytate suppresses zinc availability in monogastric animals. Zinc availability was improved by the substitution of regular soybean meal with fermented soybean meal and by the supplementation with Aspergillus niger phytase in pigs. It has been considered that phytate is easily degraded in the rumen and the availability of phytate P is high in ruminants. However, 20% of phytate in oilseed meals was not degraded in the rumen of sheep. Additionally, heating and formaldehyde treatments with oilseed meals suppressed ruminal degradation of phytate and approximately half of phytate escaped from ruminal degradation in the treated oilseed meals.     

植酸磷和植酸酶研究进展

大量研究结果显示,植酸酶在动物生产和环境保护方面具有很好的应用前景。本文从植酸磷和有效磷的关系、植酸磷测定方法、饲料中的植酸磷含量、植酸酶的来源和植酸酶在畜禽饲料中的应用等方面,就近10年来对植酸磷和植酸酶的研究动态和进展进行了简要综述。     

The effects of phytase and phytic acid on the loss of endogenous amino acids and minerals from broiler chickens

1. The effects of myo-inositol hexaphosphate (IP6) and phytase (EC 3.1.3.26) on the excretion of endogenous compounds were investigated using growing broiler chickens. 2. A total of 32 female Ross broilers were used in a precision feeding assay involving a 2 x 2 factorial arrangement of treatments. The materials administered were glucose, glucose + 1000 units of phytase activity (FTU), glucose + 1 g of IP6 and glucose + 1 g of IP6 + 1000 FTU. Excreta were collected quantitatively over a 48-h period following intubation of the test materials. The excretion of nitrogen, amino acids, minerals, sialic acid and phytate phosphorus was determined. 3. The ingestion of 1 g of IP6 by broilers increased the excretion of endogenous nitrogen, amino acids, iron, sodium, sulphur and sialic acid compared with birds fed on glucose. Supplementation of IP6 with exogenous phytase reduced the excretion of endogenous amino acids, calcium, sodium, phytate phosphorus and sialic acid compared with birds fed IP6. 4. It can be concluded that IP6 increases the excretion of endogenous minerals and amino acids in broiler chickens. Part of the beneficial effects of the addition of exogenous phytases to the diets of poultry appears to be mediated through a reduction in endogenous losses of these nutrients.     

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.