应用免疫学技术测定植酸酶活性的设想与展望

doi:10.11843/j.issn.0366-6964.2015.10.001

摘要/Abstract

摘要：

植酸广泛存在于粮食、饲料中，与磷及其他养分结合形成不溶性的络合物，降低磷和其他养分利用率。植酸酶能够分解植酸及其盐类，释放出磷元素及其他养分，因而可以在饲料中添加植酸酶来减少磷酸氢钙用量。但如果饲料中降低了磷酸氢钙添加量，而忘记添加植酸酶会造成动物缺磷，导致饲料产品质量降低，动物生产性能及产品品质下降，因而，监控检测饲料中植酸酶活性对保证饲料产品质量至关重要。目前，植酸酶常用的检测方法不适合饲料生产及畜禽养殖企业现场实时检测。本文提出了用免疫学检测技术检测植酸酶活性的设想，并展望了免疫学技术在植酸酶活性检测中应用前景。

关键词: 植酸, 植酸酶, 饲料品质, 免疫学检测

Abstract:

Phytic acid exists in food and feed widely，which can be combined with phosphorus and other nutrients to form insoluble complexes and subsequently reduce the phosphorus and other nutrient utilization rate.Phytase can decompose phytic acid and its salts to release of phosphorus and other nutrients，thus，phytase can be added in feed to replace part of dicalcium phosphate.However，if we reduce the amount of calcium hydrogen phosphate and forget to add phytase in feed in the same time，which will decline the feed quality，and subsequently lead to poor growth performance and production quality of animals.The measurement of phytase activity in feed is very important to ensure the quality of feed products.At present，the commonly detection methods are cumbersome and unsuitable for on-site detection of phytase activity for feed and animal husbandry enterprises.In this paper，a conception of detecting phytase activity with immunological techniques is proposed.Furthermore，we prospect the application of immunological techniques in the detection of phytase activity.

Key words: phytic acid, phytase, feed quality, immunoassay

中图分类号:

S816.7

胡骁飞，魏凤仙，杨继飞，邓瑞广，张改平. 应用免疫学技术测定植酸酶活性的设想与展望[J]. 畜牧兽医学报, 2015, 46(10): 1697-1705.

HU Xiao-fei，WEI Feng-xian，YANG Ji-fei，DENG Rui-guang，ZHANG Gai-ping. The Conception and Prospect of Detecting Phytase Activity by Immunological Techniques[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(10): 1697-1705.

参考文献

［1］付雪梅.植酸酶对植物性饲料植酸磷利用率影响的研究［D］.雅安：四川农业大学，2007.
FU X M.Study on the effeet of phytase on availability of phytate phosphorus in plant feed［D］.Ya’an：Sichuan Agricultural University，2007.(in Chinese)
［2］LOTT J N A，OCKENDEN I，RABOY V，et al.Phytic acid and phosphorus in crop seeds and fruits：a global estimate［J］.Seed Sci Res，2000，10(1)：11-33.
［3］NORAZALINA S，NORHAIZAN M E，HAIRUSZAH I，et al.Anticarcinogenic efficacy of phytic acid extracted from rice bran on azoxymethane-induced colon carcinogenesis in rats［J］.Exp Toxicol Pathol，2010，62(3)：259-268.
［4］ZHANG H W，BAI X L.Optimization of extraction conditions for phytic acid from rice bran using response surface methodology and its antioxidant effects［J］.J Food Sci Technol，2014，51(2)：371-376.
［5］KUNYANGA C N，IMUNGI J K，OKOTH M W，et al.Antioxidant and type 2 diabetes related functional properties of phytic acid extract from Kenyan local food ingredients：effects of traditional processing methods［J］.Ecol Food Nutr，2011，50(5)：452-471.
［6］LANDONI M，CERINO B F，HAMAN N，et al.Low phytic acid 1 mutation in maize modifies density，starch properties，cations，and fiber contents in the seed［J］.J Agric Food Chem，2013，61(19)：4622-4630.
［7］WOYENGO T A，WEIHRAUCH D，NYACHOTI C M.Effect of dietary phytic acid on performance and nutrient uptake in the small intestine of piglets［J］.J Anim Sci，2012，90(2)：543-549.
［8］HILL B E，SUTTON A L，RICHERT B T.Effects of low-phytic acid corn，low-phytic acid soybean meal，and phytase on nutrient digestibility and excretion in growing pigs［J］.J Anim Sci，2009，87(4)：1518-1527.
［9］ONYANGO E M，ASEM E K，ADEOLA O.Phytic acid increases mucin and endogenous amino acid losses from the gastrointestinal tract of chickens［J］.Br J Nutr，2009，101(6)：836-842.
［10］COSTA P S，NASCIMENTO A M，CLÁUDIA I L B，et al.Chromobacterium sp.from the tropics：detection and diversity of phytase activity［J］.Braz J Microbiol，2011，42(1)：84-88.
［11］OU K，CHENG Y，XING Y，et al.Phytase activity in brown rice during steeping and sprouting［J］.J Food Sci Technol，2011，48(5)：598-603.
［12］LOPEZ H W，VALLERY F，LEVRAT-VERNY M A，et al.Dietary phytic acid and wheat bran enhance mucosal phytase activity in rat small intestine［J］. J Nutr，2000，130(8)：2020-2025.
［13］WU T H，CHEN C C，CHENG Y S，et al.Improving specific activity and thermostability of Escherichia coli phytase by structure-based rational design［J］.J Biotechnol，2014，175：1-6.
［14］ZENG Z K，WANG D，PIAO X S，et al.Effects of adding super dose phytase to the phosphorus-deficient diets of young pigs on growth performance，bone quality，minerals and amino acids digestibilities［J］.Asian-Australas J Anim Sci，2014，27(2)：237-246.
［15］GAO C，MA Q，ZHAO L，et al.Effect of dietary phytase transgenic corn on physiological characteristics and the fate of recombinant plant DNA in laying hens［J］.Asian-Australas J Anim Sci，2014，27(1)：77-82.
［16］WALK C L，SANTOS T T，BEDFORD M R.Influence of superdoses of a novel microbial phytase on growth performance，tibia ash，and gizzard phytate and inositol in young broilers［J］.Poult Sci，2014，93(5)：1172-1177.
［17］MURUGESAN G R，ROMERO L F，PERSIA M E.Effects of protease，phytase and a Bacillus sp.direct-fed microbial on nutrient and energy digestibility，ileal brush border digestive enzyme activity and cecal short-chain fatty acid concentration in broiler chickens［J］.PLoS One，2014，9(7)：e101888.
［18］VIGORS S，SWEENEY T，O’SHEA C J，et al.Improvements in growth performance，bone mineral status and nutrient digestibility in pigs following the dietary inclusion of phytase are accompanied by modifications in intestinal nutrient transporter gene expression［J］.Br J Nutr，2014，112(5)：688-697.
［19］ZAEFARIAN F，ROMERO L F，RAVINDRAN V.Influence of a microbial phytase on the performance and the utilisation of energy，crude protein and fatty acids of young broilers fed on phosphorus-adequate maize-and wheat-based diets［J］.Br Poult Sci，2013，54(5)：653-660.
［20］BENTO M H，PEDERSEN C，PLUMSTEAD P W，et al.Dose response of a new phytase on dry matter，calcium，and phosphorus digestibility in weaned piglets［J］.J Anim Sci，2012，90(Suppl 4)：245-247.
［21］UM J S，PAIK I K.Effects of microbial phytase supplementation on egg production，eggshell quality，and mineral retention of laying hens fed different levels of phosphorus［J］.Poult Sci，1999，78(1)：75-79.
［22］BAXTER C A，JOERN B C，RAGLAND D，et al.Phytase，high-available-phosphorus corn，and storage effects on phosphorus levels in pig excreta［J］.J Environ Qual，2003，32(4)：1481-1489.
［23］RUTHERFURD S M，CHUNG T K，THOMAS D V，et al.Effect of a novel phytase on growth performance，apparent metabolizable energy，and the availability of minerals and amino acids in a low-phosphorus corn-soybean meal diet for broilers［J］.Poult Sci，2012，91(5)：1118-1127.
［24］CAREW L B J，GESTONE T A，ALSTER F A，et al.Effect of phosphorus deficiency on thyroid function and growth hormone in the white Leghorn male［J］.Poult Sci， 1985，64(10)：2010-2012.
［25］BRAUN U，OHLERTH S，LIESEGANG A，et al.Osteoporosis in goats associated with phosphorus and calcium deficiency［J］. Vet Rec，2009，164(7)：211-213.
［26］LIESEGANG A，URSPRUNG R，GASSER J，et al.Influence of dietary phosphorus deficiency with or without addition of fumaric acid to a diet in pigs on bone parameters［J］.J Anim Physiol Anim Nutr(Berl)，2002，86(1-2)：1-16.
［27］BAR A，HURWITZ S.Egg shell quality，medullary bone ash，intestinal calcium and phosphorus absorption，and calcium-binding protein in phosphorus-deficient hens［J］.Poult Sci，1984，63(10)：1975-1979.
［28］ADEMOSUN A A，KALANGO I O.Effect of calcium and phosphorus levels on the performance of layers in Nigeria.1.Egg production，egg shell quality，feed intake and body weight［J］.Poult Sci，1973，52(4)：1383-1392.
［29］HOLMAN W I.A new technique for the determination of phosphorus by the molybdenum blue method［J］.Biochem J，1943，37(2)：256-259.
［30］YOON S J，CHOI Y J，MINT H K，et al.Isolation and identification of phytase-producing bacterium，Enterobacter sp.4，and enzymatic properties of phytase enzyme［J］.Enzyme Microb Technol，1996，18：449-454.
［31］彭益强，贺淹才.几种植酸酶活测定方法的修正与差异比较［J］.福建分析测试，2002，11(2)：1564-1567.
PENG Y Q，HE Y C.Modify and differentia analysis of several phytase activity measure methods［J］.Fujian Analysis and Testing，2002，11(2)：1564-1567.(in Chinese)
［32］HEINONEN J K，LAHTI R J.A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase［J］.Anal Biochem，1981，113(2)：313-317.
［33］KIM T W，LEI X G.An improved method for a rapid determination of phytase activity in animal feed［J］.J Anim Sci，2005，83(5)：1062-1067.
［34］ENGELEN A J，VAN DER HEEFT F C，RANDSDORP P H，et al.Simple and rapid determination of phytase activity［J］. J AOAC Int，1994，77(3)：760-764.
［35］ENGELEN A J，VAN DER HEEFT F C，RANDSDORP P H，et al.Determination of phytase activity in feed by a colorimetric enzymatic method：collaborative interlaboratory study［J］. J AOAC Int，2001，84(3)：629-633.
［36］饲用植酸酶活性的测定［S］.GB/T18634-2002.Determination of phytase activity in feed［S］.GB/T18634-2002.(in Chinese)
［37］饲用植酸酶活性的测定［S］.GB/T18634-2009.Determination of phytase activity in feed［S］.GB/T18634-2009.(in Chinese)
［38］SHIEH T R，WARE J H.Survey of microorganism for the production of extracellular phytase［J］.Appl Microbiol，1968，16(9)：1348-1351.
［39］BAE H D，YANKE L J，CHENG K J，et al.A novel staining method for detecting phytase activity［J］.J Microbiol Methods，1999，39(1)：17-22.
［40］KORNBERG A，RAO N N，AULT-RICHÉ D.Inorganic polyphosphate：a molecule of many functions［J］.Annu Rev Biochem，1999，68：89-125.
［41］BERRY D F，BERRY D A.Tethered phytic acid as a probe for measuring phytase activity［J］.Bioorg Med Chem Lett， 2005，15(12)：3157-3161.
［42］杨海锋，赵志辉，秦玉昌.植酸酶酶活的近红外光谱快速检测研究［J］.中国饲料，2008(7)：36-38.
YANG H F，ZHAO Z H，QIN Y C.The study of near infrared spectroscopy for rapid detection of phytase activity［J］.China Feed，2008(7)：36-38.(in Chinese)
［43］朱立平，陈学清.免疫学常用实验方法［M］.北京：人民军医出版社，2000：467.
ZHU L P，CHEN X Q.Experimental methods in immunology［M］.Beijing：People’s Military Surgeon Press，2000：467.(in Chinese)
［44］HECKEL K，KIEFMANN R，DÖRGER M，et al.Colloidal gold particles as a new in vivo marker of early acute lung injury［J］.Am J Physiol Lung Cell Mol Physiol，2004，287(4)：L867-L878.
［45］CHEN Y，WANG Z，WANG Z，et al.Rapid enzyme-linked immunosorbent assay and colloidal gold immunoassay for kanamycin and tobramycin in swine tissues［J］.J Agric Food Chem，2008，56(9)：2944-2952.
［46］PEI S C，LEE W J，ZHANG G P，et al.Development of anti-zearalenone monoclonal antibody and detection of zearalenone in corn products from China by ELISA［J］.Food Control，2013，31（1）：65-70.
［47］ZHAO Y，ZHANG G，LIU Q，et al.Development of a lateral flow colloidal gold immunoassay strip for the rapid detection of enrofloxacin residues［J］.J Agric Food Chem，2008，56(24)：12138-12142.
［48］SONG C，LIU Q，ZHI A，et al.Development of a lateral flow colloidal gold immunoassay strip for the rapid detection of olaquindox residues［J］.J Agric Food Chem，2011，59(17)：9319-9326.
［49］ZHANG G P，WANG X N，YANG J F，et al.Development of an immunochromatographic lateral flow test strip for detection of beta-adrenergic agonist Clenbuterol residues［J］.J Immunol Methods，2006，312(1-2)：27-33.
［50］ZHANG G P，LI Q M，YANG Y Y，et al.Development of a one-step strip test for the diagnosis of chicken infectious bursal disease［J］.Avian Dis，2005，49(2)：177-181.
［51］ZHANG G P，GUO J Q，WANG X N，et al.Development and evaluation of an immunochromatographic strip for trichinellosis detection［J］.Vet Parasitol，2006，137(3-4)：286-293.
［52］胡骁飞，魏凤仙，邢广旭，等.植酸酶鼠源多克隆抗血清的制备［J］.中国农业科学，2011，44(3)：613-619.
HU X F，WEI F X，XING G X，et al.Preparation of phytase mouse polyclonal antiserum［J］.Scientia Agricultura Sinica，2011，44(3)：613-619.(in Chinese)
［53］胡骁飞，魏凤仙，李青梅，等.植酸酶单克隆抗体的制备及鉴定［J］.华北农学报，2012，27(5)：44-48.
HU X F，WEI F X，LI Q M，et al.Preparation and characteristic of monoclonal antibody against phytase［J］.Acta Agriculturae Boreali-Sinica，2012，27(5)：44-48.(in Chinese)
［54］RODRIGUEZ E，HAN Y，LEI X G.Cloning，sequencing，and expression of an Escherichia coli acid phosphatase/phytase gene(appA2) isolated from pig colon［J］.Biochem Biophys Res Commun，1999，257(1)：117-123.
［55］王爱萍，李发弟，胡骁飞，等.新霉素阻断ELISA试剂盒的研制与应用［J］.畜牧兽医学报，2011，42(6)：857-864.
WANG A P，LI F D，HU X F，et al.Development and evaluation of blocking ELISA kit for Neomycin detection［J］.Acta Veterinaria et Zootechnica Sinica，2011，42(6)：857-864.(in Chinese)
［56］YANG M，KOSTOV Y，BRUCK H A，et al.Carbon nanotubes with enhanced chemiluminescence immunoassay for CCD-based detection of Staphylococcal enterotoxin B in food［J］.Anal Chem，2008，80(22)：8532-8537.
［57］SONG C，ZHI A，LIU Q，et al.Rapid and sensitive detection of β-agonists using a portable fluorescence biosensor based on fluorescent nanosilica and a lateral flow test strip［J］.Biosens Bioelectron，2013，50：62-65.

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