木聚糖酶TAXI型抑制剂及基因工程研究

谷物中存在2种类型的木聚糖酶抑制剂,根据其结构的不同可分为TAXI型和XIP型。本文主要简述了木聚糖酶TAXI型抑制剂对木聚糖酶的抑制特性及其基因工程研究。     

木聚糖酶抑制蛋白对木聚糖酶应用领域的影响

木聚糖酶已广泛应用于饲料、食品加工、纸浆漂白等领域,然而近年研究发现在小麦等谷物中存在一种能抑制木聚糖酶活性的蛋白质性质的成分,称为木聚糖酶抑制蛋白.木聚糖酶抑制蛋白具有多型性,但不同类型抑制蛋白都只作用于外源木聚糖酶,而对谷物内源性木聚糖酶没有抑制作用.这就对木聚糖酶应用领域中酶功效的发挥提出了挑战:抑制蛋白的存在是...     

固态发酵木聚糖酶的研究进展

木聚糖酶(xylanase)是指将木聚糖降解成低聚糖和木糖的一组酶的总称,是木聚糖降解酶系中最关键的酶,在饲料、造纸、食品和生物转化等行业应用广泛。文中对固态发酵木聚糖酶的微生物菌种、诱导底物以及主要影响因素等方面进行了综述。     

饲用木聚糖酶及其抑制蛋白研究进展综述

木聚糖酶是半纤维素酶类的其中一种,它能水解可溶性非淀粉多糖,摧毁植物性饲料的细胞壁,降低食糜黏度,提高谷物饲料利用率,对利用谷物充当能量饲料具有重要意义。本文对木聚糖酶及其抑制蛋白的作用机理进行综述,以期为今后木聚糖酶制剂的开发应用提供参考。     

饲用木聚糖酶制剂的体外评价技术研究

试验旨在研究木聚糖酶对蛋白酶的耐受性、对饲料还原糖含量及饲料能量和粗蛋白消化率的影响,以期建立木聚糖酶体外评价的方法。采集不同厂家的5种木聚糖酶制剂,经胃蛋白酶和胰蛋白酶两步反应后,测定木聚糖酶的酶活留存率,以一定的比例在猪配合饲料中添加木聚糖酶,保证饲料中酶活性相同,采用仿生消化系统测定粗蛋白和能量的消化率,同时模拟胃肠道p H值测定饲料中还原糖含量。结果表明:通过胃蛋白酶和胰蛋白酶的孵育后,5种木聚糖酶留存率差异显著(P0.05),说明不同木聚糖酶对蛋白酶的耐受性不同。但饲料中添加5种不同的木聚糖酶制剂,对饲料能量和粗蛋白消化率及还原糖含量均没有显著影响(P0.05)。因此,在试验条件下,两步酶解法对木聚糖酶留存率影响显著,可以作为木聚糖酶质量评价的参考;但是采用仿生消化系统测定饲料中能量和粗蛋白的消化率以及还原糖的含量,来评价木聚糖酶的生物学效应,还需要进一步通过动物试验进行验证。     

微生物发酵木聚糖酶的研究进展

木聚糖酶（xylanase）是指可将木聚糖降解成低聚糖和木糖的一组酶的总称,主要包括外切／3—1,4-木聚糖酶、内切口-1,4-木聚糖酶和／3-木聚糖酶。木聚糖酶在饲料、食品、造纸、纺织、医药及能源等领域有着广泛的应用。作为饲用酶制剂,木聚糖酶可以有效解除木聚糖的抗营养作用,促进畜禽对粗饲料的消化吸收;作为新型的纸浆漂...     

钙蛋白酶抑制蛋白与肉嫩度相关性

钙蛋白酶蛋白系统是影响肌肉生长和宰后肉嫩化的一个重要因素,主要由钙蛋白酶(μ-calpain和m-cal-pain)、钙蛋白酶抑制蛋白(calpastatin,CAST)及骨骼肌特异性钙蛋白酶(muscle specific calpain,P94)组成,钙蛋白酶抑制蛋白是一种内源性专一抑制钙蛋白酶(calpain)活性的蛋白,参与了调控肌原纤维蛋白的降解并在其中发挥了关键作用。本文概述了钙蛋白酶系统、钙蛋白酶抑制蛋白的结构、功能及其对提高肉嫩度的作用。     

离子注入选育高产木聚糖酶黑曲霉及其发酵条件研究

木聚糖酶是一类能够特异降解木聚糖的酶类,其水解产物以低聚木糖为主,并伴有少量木糖和阿拉伯糖。研究表明,饲料中含有的非淀粉多糖(NSP)是饲料中重要的抗营养因子,木聚糖是其中之一。在饲料中添加木聚糖酶可消除木聚糖的抗营养作用,通过降低肠道内容物的黏度,提高动物内源性消化酶的活性,从而提高饲料转化率,促进养分的消化吸收。木聚糖酶酶解产生的低聚木糖有利于促进肠道内健康菌的形成,刺激动物机体的免疫反应机能,抑制病原菌的生长,减少疾病(陈瑞娟,1 993)。木聚糖酶与其他非淀粉多糖酶(如纤维素酶、β-葡聚糖酶、果胶酶)复合使用作…     

钙蛋白酶抑制蛋白对猪肉品质的影响

钙蛋白酶蛋白系统是影响肌肉生长和宰后肉嫩化的一个重要因素,主要由钙蛋白酶(μ-calpain和m-calpain)、钙蛋白酶抑制蛋白(calpastatin,CAST)及骨骼肌特异性钙蛋白酶组成,钙蛋白酶抑制蛋白是一种内源性专一抑制钙蛋白酶活性的蛋白,参与了调控肌原纤维蛋白的降解并在其中发挥了关键作用。本文概述了钙蛋白酶抑制蛋白的结构、功能及其对肌肉蛋白质的增加、提高肉嫩度的作用。     

MBTH法检测木聚糖酶活性

试验采用3-甲基-2-苯并噻唑酮腙(MBTH)法检测饲料添加剂木聚糖酶和饲料中木聚糖酶的活性。试验结果表明,MBTH法既可用于检测木聚糖酶制剂的活性,又可用于饲料中木聚糖酶的检测,但若能更好地降低饲料本身对样品空白值的干扰则更能体现该方法的优势。     

Effect of cereal type and Xylanase supplementation on nutrient retention and growth performance of broilers

An experiment was conducted to study the effect of cereal (corn, wheat, barley and sorghum) and supplemental enzyme (a mono-component xylanase) in a 4 × 2 factorial arrangement. A 2-phase feeding program (0 to 21 and 21 to 42 d) was used and 4 iso-caloric, iso-nitrogenous diets were formulated containing corn, wheat, barley, or sorghum as the sole cereal. Diets based on each cereal were fed without or with supplemental xylanase (16,000 BXU/kg) to 8 replicate pens of 10 chicks (5 male : 5 female, Cobb 500) each. Growth performance was recorded at 21 or 42 d posthatch. Excreta was quantitatively collected from 18 to 21 and 38 to 41 d for the measurement of the total tract retention of dry matter (DM), nitrogen (N) and neutral detergent fiber (NDF) and apparent metabolizable energy (AME). Ileal digestible energy (IDE) was measured at the end of the study (42 d posthatch) using titanium dioxide as an indigestible marker. For the overall 42 d period, birds fed barley-based diets had lower feed intake (P < 0.05), lower body weight (P < 0.05) and converted feed into gain less efficiently (P < 0.05) compared with the diets based on corn, wheat or sorghum. Xylanase supplementation improved weight gain in diets based on corn, wheat, and sorghum with the exception of barley-based diets (cereal × xylanase interaction, P < 0.05). Xylanase improved the overall feed conversion ratio (1.885 vs. 1.939; P < 0.05) with the effect being independent of the cereal type. The N retention of barley-based diets was lower (P < 0.05) compared with the other cereals, while xylanase improved N retention (P < 0.05) regardless of the cereal type. NDF digestibility differed (P < 0.05) across cereal (barley > wheat > corn > sorghum) and was improved (P < 0.05) by xylanase supplementation. A significant cereal × xylanase interaction (P < 0.05) was observed for energy measurements, where xylanase improved IDE of the corn-based diet, and AME of corn- and wheat-based diets. Results of the current study demonstrate potential of xylanase in improving nutrient retention and growth performance of broilers fed diets based on variety of cereal grains.     

Xylanase Inhibitors Affect the Action of Exogenous Enzymes Used to Supplement Triticum durum-Based Diets for Broiler Chicks

Diets containing low-quality durum wheat (716 g/kg) for production of pasta were used to feed male broiler chicks. The efficacy of xylanase supplementation and the impact of xylanase inhibitors on losses in exogenous enzyme activity were analyzed. Birds fed on the basal diet, not supplemented with recombinant xylanases or Roxazyme G, reached a BW of 1,509 g with a feed conversion ratio of 1.77 at d 28. Growing performance was above that expected for the breed used, whereas feed conversion ratios were relatively higher. None of the 3 xylanase preparations under analysis affected growing performances and feed efficiency of broiler chicks. The activity of feed xylanases was considerably reduced in the presence of durum wheat extracts. The results suggest that reduction of exogenous enzyme activity was due to the action of durum wheat xylanase inhibitors and not to proteolysis.     

Bacterial responses to different dietary cereal types and xylanase supplementation in the intestine of broiler chicken.

Several studies were carried out to investigate the influence of dietary cereals differing in soluble non starch polysaccharides (NSP) content and a xylanase preparation on selected bacterial parameters in the small intestine of broiler chicken. Compared to a maize diet colony forming units (CFU) of mucosa associated bacteria were higher in a wheat/rye diet, most notably for enterobacteria and enterococci. Xylanase supplementation to the wheat/rye diet generally led to lower CFU, especially in the first week of life. However, xylanase supplementation also displayed higher in vitro growth potentials for enterobacteria and enterococci. Bacterial growth of luminal samples in minimal media supplemented with selected NSP showed that the wheat/rye diet enhanced bacterial capacities to utilize NSP only in ileal samples. The xylanase application generally shifted respective maximum growth to the proximal part of the small intestine. The presence of soluble NSP from wheat or rye in the diet per se did not enhance bacterial NSP hydrolyzing enzyme activities in the small intestine, but xylanase supplementation resulted in higher 1,3-1,4-beta-glucanase activity. Compared to a maize diet the activity of bacterial bile salt hydrolases in samples of the small intestine was not increased due to inclusion of wheat/rye or triticale to the diet. However, xylanase supplementation led to a reduction with a corresponding increase of lipase activity. It was concluded that dietary cereals producing high intestinal viscosities lead to increased overall bacterial activity in the small intestine. The supplementation of a xylanase to cereal based diets producing high intestinal viscosity, changes composition and metabolic potential of bacterial populations and may specifically influence fat absorption in young animals.     

Beta-glucanase and xylanase activities in stomach and ileum of growing pigs fed wheat bran based diets with and without enzyme treatment.

Pigs were fed diets containing 40% wheat bran incubated with a water: acetic acid mixture (control C) and a cellulase (Cel-i) or xylanase (Xyl-i) preparation or with addition of the cellulase (Cel-a) or xylanase (Xyl-a) preparation immediately before feeding. Stomach and ileal samples were analysed for xylanase and beta-glucanase activities by a standard method based on release of reducing substances and a modified method in which the actual pH of the sample was used. After feeding Cel-treated diets xylanase and beta-glucanase activities were higher in the stomach up to 4 hours after feeding. In the stomach the modified method gave significantly lower xylanase activities than the standard method. Ileal xylanase activity was significantly higher 2-6 hours after feeding Xyl-treated diets and ileal beta-glucanase activity was higher 4-6 hours after feeding Cel-treated diets. In ileal samples, the modified method gave significantly higher xylanase and lower beta-glucanase activities than the standard method. It is concluded that xylanase and beta-glucanase activities can be measured in digesta samples with the methods employed and that the added enzymes were largely inactivated between the feedings. Measuring digesta enzyme activities in conditions (pH) similar to the ones in the gastrointestinal tract may give a more reliable picture of the actual activity of added enzymes.     

Characterisation and effects of a xylanase enzyme preparation extracted from Thermomyces lanuginosus cultures

This paper describes the production of an enzyme preparation from the fungus Thermomyces lanuginosus. Thermal resistance, pH stability and lignocellulolytic activity of the enzyme preparation high in xylanase were studied on a variety of grains and forages. The enzyme preparation preserved more than 70% of its original xylanase activity for 4 and 1 h at 60 and 70 degrees C, respectively. The xylanase activity remained over 80% when the preparation was incubated for 30 min at pH 4.5. In vitro digestibility studies indicated that the enzyme digested 7.5, 8.5 and 8.0% of the dry matter (DM) of barley meal, wheat bran and oat meal samples, respectively. When applying 60-min incubation, 7.5, 7.3 and 8.4% of DM of the oat straw, alfalfa hay and triticale straw was digested, respectively. When the time of digestion was increased to 360 min, the sunflower hull showed 15.8% DM digestibility.     

Effect of β -glucanase and xylanase supplementation of barley- and rye-based diets on caecal microbiota of broiler chickens

1. The aim was to investigate the effect of grain type (barley or rye) and exogenous enzymes (β-glucanase or xylanase) on the composition of chicken caecal microbiota as examined by classical culturing and molecular techniques (fluorescent in-situ hybridisation (FISH) and terminal-restriction fragment-length polymorphism (T-RFLP)). 2. Plate counting revealed higher total numbers of anaerobic bacteria, lactic acid bacteria and yeasts in caecal contents of birds fed with rye-based diets than in birds fed with barley-based diets. 3. As assessed by FISH analysis, the most abundant bacterial groups in the broiler caeca were Clostridium coccoides-Eubacterium rectale followed by Bacteroides sp., Lactobacillus sp./Enterococcus sp., Bifidobacterium sp. and Enterobacteriaceae. For both cereal types, the enzyme supplementation significantly decreased the relative amount of Enterobacteriaceae. 4. The T-RFLP profiles indicated that the caecal microbiota of birds receiving rye-based diets was more diverse than that of birds fed on barley-based diets. 5. Irrespective of the method applied, the results indicate that the cereal type as well as the exogenous enzyme supplementation influence the microbiota in broiler chicken caeca, and may have the effect of reducing potentially pathogenic Enterobacteriaceae populations.     

Feed endoxylanase type and dose affect arabinoxylan hydrolysis and fermentation in ageing broilers

Despite the general use of endoxylanases in poultry feed to improve broiler performance, the abundance of different endoxylanase products and the variable response to their application in the field prevent a clear understanding of endoxylanase functionality in vivo. To gain insight into this functionality, we investigated the impact of endoxylanase type (Belfeed from Bacillus subtilis versus Econase XT from Nonomuraea flexuosa) and dose (10, 100, 1,000 mg/kg) in combination with broiler age on arabinoxylan (AX) hydrolysis and fermentation in broilers (Ross 308) fed a wheat-soy based diet. In a digestibility trial and a performance trial, a total of 1,057 one-day-old chicks received the control diet or 1 of the 6 endoxylanase supplemented wheat-soy based diets with, respectively, 5 replicate cages and 8 replicate pens per dietary treatment per trial. The AX content and structure, the AX digestibility values and the short-chain fatty acids produced were analysed at the level of the ileum, caeca and excreta at d 11 and 36. Endoxylanase supplementation resulted in a more extensive solubilisation of wheat AX and a reduction in the intestinal viscosity compared to the control (P < 0.05). A high endoxylanase dose was, however, required to obtain increased hydrolysis of the dietary AX along the gastrointestinal tract against the control (P < 0.001). Depending on the type of endoxylanase, a pool of AX with distinct physicochemical properties was created. The B. subtilis endoxylanase created a large pool of soluble AX in the ileum, thereby increasing ileal viscosity compared to broilers fed an endoxylanase from N. flexuosa (P < 0.001). The N. flexuosa endoxylanase mainly triggered caecal AX fermentation in young broilers, by delivering easily fermentable AX substrates with a low degree of polymerisation (P = 0.03). The effects were particularly present in young broilers (d 11). From this study, it is clear that the type and dose of endoxylanase added to wheat-soy based diets determine the nature of AX substrates formed. These, in turn, affect the intestinal viscosity and the interplay between the dietary AX compounds and microbiota, hence dictating AX digestion at young broiler ages and performance outcomes towards slaughter age.     

4种芽孢杆菌来源木聚糖酶基因在大肠杆菌中的表达及酶活分析

研究旨在比较4种不同来源木聚糖酶基因在大肠杆菌E.coli BL21中表达后的酶活等性质,为生产中筛选高表达量及耐极性木聚糖酶基因提供依据。根据已发表的木聚糖酶基因序列设计引物,分别扩增出浸麻芽孢杆菌(B.macerans)B3、地衣芽孢杆菌(B.licheniformis)B6、蜡样芽孢杆菌(B.cereus)B10和枯草芽孢杆菌(B.subtilis)B12的木聚糖酶基因片段,经克隆表达后获得4种工程菌。经培养和IPTG诱导后进行SDS-PAGE电泳检测及酶特性分析。测序结果表明,4个基因的开放阅读框均为642 bp。其中前3种木聚糖酶基因均为首次报道。4种重组木聚糖酶在55℃下的酶活分别为:36.16、3.85、7.22、98.98 U/ml。4种重组木聚糖酶的最适反应温度均在50~60℃,且枯草芽孢杆菌木聚糖酶最为耐热。结果提示,枯草芽孢杆菌来源的木聚糖酶具有较高的酶活和较好的耐热性。     

Effects of yeast culture and fibrolytic enzyme supplementation on in vitro fermentation characteristics of low-quality cereal straws

The effects of yeast culture and fibrolytic enzyme preparation (containing cellulase and xylanase) on in vitro fermentation characteristics of rice straw, wheat straw, maize stover, and maize stover silage were examined using an in vitro gas production technique. Four levels of yeast culture and fibrolytic enzyme supplements (0, 2.5, 5.0, and 7.5 g/kg of straw DM, respectively) were tested in a 4 x 4 factorial arrangement. Supplementation of yeast culture increased the cumulative gas production, theoretical maximum of gas production, rate of gas production, IVDMD, and in vitro OM disappearance (IVOMD), and decreased the lag time for each type of straw. Fibrolytic enzyme supplementation tended to increase cumulative gas production, theoretical maximum of gas production, and rate of gas production; prolonged lag time of gas production; and enhanced IVDMD and IVOMD for 4 types of cereal straws, with the significance of this effect being dependent on the level of supplemented enzymes. There were significant interactions between fibrolytic enzymes and yeast on all in vitro gas production parameters, IVDMD, and IVOMD of each type of straw. The outcome of this research indicated that the application of fibrolytic enzyme preparation and yeast culture could improve in vitro gas production fermentation of cereal straws.