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Enzymology properties of two different xylanases and their impacts on growth performance and intestinal microflora of weaned piglets

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Animal Nutrition 2016年第1期2卷 18-23页

作者： Qinghua Chen Mengwei Li Xiong Wang College of Animal Science and Technology Hunan Agricultural University

The enzyme xylanase is more and more widely used in feed production, but different xylanase have different properties, mechanism and application effects. To provide a theoretical basis for choosing more suitable xylanase in feed production, we selected bacterial xylanase(BX), labeled enzyme A, and trichoderma xylanase(TX), labeled enzyme B, and studied the enzymology properties and application effects on growth performance and gut flora in weaned piglets. The results showed that the activity levels of both appear parabolic along with increasing pH or temperature, but the amplitude of enzyme activity changing curves and the pH/temperature of optimal activity level are different, where enzyme A has the optimal activity level at 50 ℃ with a pH value of 5.0. The optimal activity level of enzyme B was achieved at 70 ℃ with a pH around 6.0. Enzyme B suffered very little activity loss with moisture level at16% and temperature from 80℃ to 90 ℃. Enzyme A suffered a big drop in activity level when processed with high temperature from around 80 ℃ to 90 ℃, and it was even completely inactivated at 90 ℃,Enzyme A has very low activity level after being processed in acid environment, but enzyme B has minor changes in activity level with respect to changes in acid level, indicating significantly different enzymatic properties between the two different sources of xylanases. In feeding experiment, the control group, was fed the basal diet, and the BX group and TX group were fed basal diets supplemented with 0,01% bacterial and fungal xylanases, respectively. The results showed that ADG of the BX group and TX group increased by 3.25%(P> 0.05) and 8.22%(P 0.05), respectively compared with the control group; TX group had significantly higher(P < 0.05) ADG compared with BX group; BX group and TX group had significantly lower ileum Escherichia coli level than the control group, which were reduced by up t

关键词： xylanase Enzymology characteristics Weaned piglet Growth performance Microbial flora

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Eukaryotic Secrete fusion expression of Rhizopus oryzae glucoamylase and Paenibacillus ***

Eukaryotic Secrete fusion expression of Rhizopus oryzae gluc...

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第五届全国微生物资源学术暨国家微生物资源平台运行服务研讨会

作者： Wang Zhi Dun Bao-Qing Lu Ming The National Key Facility for Crop Gene Resources and Genetic Improvement of Institute of Crop Sciences Chinese Academy of Agricultural Sciences

The glucoamylase gene(amy A) was amplified from Rhizopus oryzae 3.866 total DNA extracts. The result showed that the sequence of amyA was consisted of 2049 bp,and encoded 604 amino *** xylanase gene(xynA) was amplified from Paenibacillus sp.H10-3 total DNA *** result showed that the mature peptide sequence of xynA was consisted of 636bp,and encoded 211 amino ***,the mature peptide sequence of amyA gene,sequence of linker and sequence of xyn A gene were spliced by overlap extension PCR(SOE-PCR),which amyA-l-xynA was *** fusion gene was then inserted into a Pichia pastoris expression vector pPIC9 to produce the recombinant expression plasmid p?*** using electric shocks,we successfully transformed the recombinant pP\C9-amyA-l-xynA into Pichia pastoris GS115,which AX 11 was obtained..The maximum yield of the recombinant glucoamylase and xylanase in AX11 culture medium were 5.8U/mL and 32.3U/mL, respectively.

关键词： glucoamylase xylanase fusion expression Pichia pastoris

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Non-starch polysaccharide-degrading enzymes may improve performance when included in wheat- but not maize-based diets fed to broiler chickens under subclinical necrotic enteritis challenge

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Animal Nutrition 2022年第3期10卷 54-67页

作者： Eunjoo Kim Amy F.Moss Natalie K.Morgan Kosar Gharib-Naseri Peter Ader Mingan Choct School of Environmental and Rural Science University of New EnglandArmidaleNSW 2351Australia BASF SE 68623LampertheimGermany

The present study investigated whether supplementing fibre-degrading enzymes can ameliorate the severity of subclinical necrotic enteritis(NE)in broiler chickens offered wheat-or maize-based diets.A total of 1,544 mixed-sex broiler chickens were assigned to 16 experimental treatments as a 2×2×4 factorial arrangement of *** factors were the following:NE challenge,yes or no;diet type,wheat-or maize-based;and enzyme supplementation,control(no enzyme),family 10 xylanase(XYN10),family 11 xylanase(XYN11)or b-mannanase(MAN).Each treatment was replicated 6 times,with 16 birds per replicate pen.A three-way challenge×diet type×enzyme interaction occurred for body weight at 21 d of age(P=0.025)and overall feed conversion ratio(P=0.001).In the non-challenged birds fed the wheat-based diet,supplementing MAN increased d 21 body weight compared to the *** challenged birds fed the maize-based diet,supplemental XYN11 impeded body weight and overall FCR compared to the *** offered the maize-based diet presented heavier relative gizzard weights at both 16 and 21 d of age(P<0.001)and reduced liveability(P=0.046)compared to those fed the wheatbased *** supplementation reduced ileal and jejunal digesta viscosity at 16 d of age only in birds fed the wheat-based diet(P<0.001).XYN11 increased ileal digesta viscosity in birds fed the maizebased diet,and MAN reduced it in birds fed the wheat-based diet at 21 d of age(P=0.030).Supplementing XYN11 improved ileal soluble non-starch polysaccharides(NSP)digestibility in birds fed the wheat-based diet compared to non-supplemented birds(P<0.001).Birds fed the wheat-based diet displayed a higher abundance of Bifidobacterium,Lactobacillus and Enterobacteriaceae and butyric acid in the caeca at 16 d of age compared to birds fed the maize-based diet(P<0.05).In conclusion,supplemental XYN11 exacerbated the negative impact of NE on growth performance in birds fed the maizebased *** wheat-based diets with

关键词： Broiler chicken Carbohydrase Mannanase Necrotic enteritis Prebiotic oligosaccharides xylanase

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Large intestinal dynamics differ between fowl and swine:Anatomical modifications,microbial collaboration,and digestive advantages from fibrolytic enzymes

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Animal Nutrition 2022年第4期11卷 160-170页

作者： Edwin T.Moran Jr. Michael R.Bedford Poultry Science Department Auburn UniversityAL 36830-5416USA AB Vista Woodstock CourtBlenheim RoadMarlboroughWiltshire SN84ANUK

The large intestinal systems of fowl and swine recover nutrients from ileal indigesta by a strategically different *** with fowl enter a short colon where retro-peristalsis using urine from the urodeum carries small particulates and solutes into both ceca while coarse materials collect in the *** repetitively add fine and soluble materials into both ceca to continue fermentation until complexity of the remainder exceeds microbial action,then contents apart from faeces are entirely *** with swine initially enter a short cecum followed by a lengthy progression through to the rectal *** out-pocketings of circular muscle or haustrae occur throughout the length of the pig's cecum and helicoidal *** pocket carries contents acquired earlier in the *** collects fines and solutes into haustrae during their progression through the colon whereas coarse particulates assemble in the *** contents continually ferment during movement to the distal colon with resulting volatile fatty acids(VFA)and electrolytes being *** loosely covers the lumen surface in caeca as well as helicoidal colon that may capture microbes from active intestinal contents as well as release others to sustain *** microbial community continually modifies to accommodate fibre complexity as *** starches(RS)and simple oligosaccharides rapidly ferment to yield VFA while encouraging butyric acid in the cecum and anterior colon,whereas non-starch polysaccharides(NSP)complexity requires extended durations through the remaining colon that enhance acetic *** fibre eventually results in undue complexity for fermentation and consolidates at termination of the *** compact pellets are placed on core contents to form faeces having a nodular ***,propionic,and butyric acids represent the bulk of VFA and are derived from non-digestible *** enzymes,when supplemented

关键词： Butyric acid Endogenous protein Intestinal microbes Large intestine Volatile fatty acids xylanase

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Screening of a Composite Microbial System and Its Characteristics of Wheat Straw Degradation

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Agricultural Sciences in China 2011年第10期10卷 1586-1594页

作者： LI Pei-pei WANG Xiao-juan YUAN Xu-feng WANG Xiao-fen CAO Yan-zuan CUI Zong-jun College of Agronomy and Biotechnology China Agricultural University Beijing 100193 P.R.China

To accelerate the decomposition of wheat straw directly returned to soil, we constructed a microbial system （ADS-3） from agricultural soil containing rotting straw residues using a 40-wk limited cultivation. To assess its potential use for accelerating straw decomposing, the decomposing characteristics and the microbial composition of ADS-3 were analyzed. The results indicated that it could degrade wheat straw and filter paper by 63.8 and 80%, respectively, during 15 d of incubation. Straw hemicellulose degraded dramatically 51.2% during the first 3 d, decreasing up to 73.7% by the end of incubation. Cellulose showed sustained degradation reaching 53.3% in 15 d. Peak values of xylanase and cellulase activities appeared at 3 and 11 d, with 1.32 and 0.15 U mL-1, respectively. Estimated pH averaged 6.4-7.6 during the degradation process, which approximated acidity and alkalinity of normal soils. The microbial composition of ADS-3 was stable based on denaturing gradient gel electrophoresis （DGGE） analysis. By using bacterial 16S rRNA and fungal 26S rRNA gene clone library analysis, 20 bacterial clones and 7 fungal clones were detected. Closest identified relatives of bacteria represented by Bacillus fusiformis, Cytophaga sp., uncultured Clostridiales bacterium, Ruminobacillus xylanolyticum, Clostridium hydroxybenzoicum, and uncultured proteobacterium and the fungi were mainly identified as related to Pichia sp. and uncultured fungus.

关键词： wheat straw degradation composite microbial system cellulose hemicelulose cellulase xylanase microbial composition

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Improving sorghum digestion in broilers by targeting fermentation of xylan

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Animal Nutrition 2022年第3期10卷 198-206页

作者： Natalie K.Morgan Andrew Wallace Michael R.Bedford University of New England School of Environmental and Rural ScienceArmidaleNew South WalesAustralia AB Vista Woodstock CourtBlenheim RoadMarlborough Business ParkMarlboroughWiltshireEnglandUnited Kingdom Curtin University Kent StreetBentleyWestern AustraliaAustralia

This study was to examine if it is possible to accelerate sorghum digestion in broiler chickens by targeting fermentation of the *** 500 broilers(n=960,80 birds per treatment)were fed 12 sorghumsoybean meal-based dietary treatments fed as 3 phases(starter d 0 to 12,grower d 13 to 23,finisher d 24 to 35),with 8 replicate pens of 10 birds per *** half of the treatments(n=6),10%of the sorghum in the diet was directly replaced with 10%wheat bran,as a source of fermentable *** diets were supplemented with either 0,50 or 2,000 mg/kg xylo-oligosaccharides(XOS),with or without xylanase *** weight gain(BWG),feed intake(FI)and feed conversion corrected for mortality(cFCR)was determined at d 0 to 35,and male and female body weight were measured on d *** d 35,ileum and caeca samples were collected from 2 birds per pen,for determination of caecal cellulase and xylanase activity,microbiota composition and short chain fatty acid(SCFA)concentration,and ileal XOS *** with 2,000 mg/kg XOS caused increased BWG at d 0 to 35(P=0.007)and enhanced caecal propionic,valeric and succinic acid concentration(Pxylanase,resulting in increased caecal total SCFA,acetic acid and butyric acid concentration,and xylanase and cellulase activity(Pxylanase and XOS.

关键词： xylanase Sorghum Oligosaccharides Broiler

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Identification of proteins involved in lignocellulose degradation using in gel zymogram analysis combined with mass spectroscopy-based peptide analysis of gut proteins from larval Asian Ionghorned beetles, Anoplophora glabripennis

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Insect Science 2010年第3期17卷 253-264页

作者： Scott M. Geib Ming Tien Kelli Hoover Departments of Biochemistry and Molecular Biology Departments of Entomology The Pennsylvania State University University Park Pennsylvania USA

Enzyme activities toward lignocellulose substrates were analyzed in the gut of larval Asian longhorned beetle （Anoplophora glabripennis）. Total protein was extracted from gut contents of wild collected larvae from an invasive population in Worchester, MA, USA. From these protein extracts, lignocellulolytic activities were measured （β-1,4- endoglucanase,β-1,4-glucosidase and birch wood xylanase）.β-1,4-glucosidase activity was 0.075 μmol glucose/mg protein per min, endoglucanase activity was measured at 0.41/zmol glucose/mg protein per min and xylanase activity was 0.058μmol xylose/mg protein per min. To identify specific enzymes that may provide these activities, zymogram analysis was performed to detect enzymes active toward carboxymethyl cellulose （CMC）, 4-methylumbelliferyl-β-D-glueopyranoside and birch wood xylan. Three protein bands were found to be active toward CMC, three displayed β-1,4-glucosidase, and one displayed xylanase activity. Proteins from active bands from these zymograms were then identified by in-gel trypsin digestions followed by peptide identification by matrix-assisted laser desorption ionization - time of flight - time of flight mass spectrometry （MS）. A custom A. glabripennis transcriptome database was used for peptide identification, giving highly significant matches in all MS analyses. These matches were then searched against the National Center for Biotechnology Information database to provide annotation to the transcripts and provide possible classification. From these analyses, we were able to detect enzymes active toward cellulose and xylan, and proteins putatively involved in lignocellulose degradation in the gut of this wood-feeding insect. Future research will be focused on characterizing these enzymes through cloning and expression experiments and understanding how the lignocellulose degradation system functions in the gut of this insect.

关键词： beta-glucosidase cellulase Cerambycidae endoglucanase proteomics xylanase

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Effect of arabinoxylo-oligosaccharides and arabinoxylans on net energy and nutrient utilization in broilers

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Animal Nutrition 2019年第1期5卷 56-62页

作者： Natalie K.Morgan Chake Keerqin Andrew Wallace Shu-Biao Wu Mingan Choct University of New England School of Environmental and Rural Science University of New England School of Science & Technology

Arabinoxylo-oligosaccharides(AXOS) are hydrolytic degradation products of arabinoxylans(AX) that can be fermented by the gut microbiota, thus potentially displaying prebiotic properties. This study examined the effects of AX and AXOS on net energy(NE) and nutrient utilization in broilers. Ross 308 broilers(n = 90, 30 birds per treatment) were fed wheat-soybean diets supplemented with pure AX, AXOS produced by exposing the AX to xylanase in vitro(AXOS), or AX with xylanase(AX + E) from d 10 to *** parameters were measured from d 10 to 21. On d 15, 10 birds per treatment were allocated to closed-circuit net energy chambers to assess the impact of AX and AXOS on dietary energy utilization,through assessment of both metabolisable energy(ME) and NE. Ileal and caecal digesta samples were collected on d 21 to determine the effect of AX and AXOS on ileal and total tract dry matter digestibility,ileal digestible energy, digesta pH, short chain fatty acids(SCFA) and microbiota concentration. Feed conversion ratio was numerically the lowest in birds fed the diet supplemented with AXOS, which is 1.26 compared to 1.37 and 1.30 for AX and AX + E, respectively. Ileal dry matter digestibility was higher in birds fed AXOS than those fed AX(P = 0.047). Ileal digestible energy and total tract dry matter digestibility were higher in birds fed AXOS than those fed AX or AX + E(P = 0.004 and P = 0.001,respectively). Birds fed AXOS had higher ME intake(P = 0.049) and nitrogen retention(P = 0.001) and a strong trend of higher NE(P = 0.056), NE intake(P = 0.057) and retained energy(P = 0.054) compared to those fed AX. Ileal total SCFA, lactic and formic acid concentrations were higher in birds fed AXOS than those fed AX(P = 0.011, P = 0.012 and P = 0.023, respectively). Birds fed AXOS or AX + E had higher caecal total SCFA, acetic, butyric and isovaleric acid concentrations compared to those fed AX(P = 0.001,P = 0.004, P = 0.016 and P = 0.008, respectively), and caecal propionic a

关键词： Arabinoxylan Oligosaccharides xylanase Broilers Prebiotic

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Pattern of non-starch polysaccharide digestion along the gut of the pig:Contribution to available energy

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Animal Nutrition 2015年第3期1卷 160-165页

作者： David J. Cadogan Mingan Choct Feedworks Pty Ltd School of Environmental and Rural Science University of New England

We investigated the pattern of non-starch polysaccharide(NSP) digestion along the gut of pigs fed two different wheats, which were offered with or without xylanase supplementation. The two wheats used were pre-characterised before the experiment on the basis of low and normal feed intake of young *** type significantly influenced feed intake and growth rate in the first 7 days, however, by day 14 the only significant effect of wheat type was on growth rate. xylanase supplementation increased the growth performance of pigs fed the poor quality wheat to a level similar to those fed the normal wheat. It also increased the daily gain of pigs fed the normal wheat. Wheat type had no significant effect on the digestibility of dry matter(DM), energy, free sugars or the different fractions of NSP in the duodenum,ileum or in the faeces. The duodenal gross energy digestibility values for the low and high performance diets were -27.4 and -47.5%, respectively, and xylanase supplementation significantly increased the digestibility of energy back to positive levels. Dry matter digestibility values followed a similar pattern. In the duodenum, xylanase increased(P xylanase had a significant effect only on the digestibility of the soluble NSP fraction. xylanase did not affect free sugar digestibility. The reduction in soluble NSP level coincided with a marked reduction in the amount of fucose, a prominent component of mucosal polysaccharides. This suggests that soluble NSP substantially increase endogenous losses. The absence of differences in the digestibility of the measured NSP between the two wheat samples suggests that the structures of the NSP, rather than just their amount and solubility, are important for the anti-nutritional properties of NSP in pig diets.

关键词： Wheat Non-starch polysaccharides xylanase Pigs

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Effect of Indica Rice Diets Supplemented with Exogenous Enzyme on Growth and Digestion of Pigs Fed Paddy-Based Diets

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Rice Science 2004年第Z1期 336-341页

作者： WANG Min-qi, Xu Zi-rong, SUN Jian-yi (Key Laboratory of Molecular Animal Nutrition, Ministry of Education Feed Science Institute Zhejiang University, Hangzhou 310029, China)

Sixty growing pigs (Landrace x Jiaxing black) were used to study the effects of exogenous enzymes (including β-glucanase, xylanase and cellulase) supplemented to paddy-based diets on pig growth performance and digestion. With the supplementation of enzymes, average daily gain of pig was increased by 8.78%(P<0.05), feed gain radio was decreased by 9.42%(P<0.05), feed apparent digestibilities of crude protein, crude fat and crude fiber were increased by 17.32% (P<0.01), 16.04% (P<0.05) and 108.57%(P<0.01), the viscosity of jejunum, and ileum contents were reduced by 7.08%(P<0.01) and 6.78%(P<0.01), numbers of E. coli in feaces and diarrhoea morbidity were decreased by 81.29%(P<0.01) and 37.00%(P<0.05), and the activities of protelytic enzyme and a-amylase in duodenal contents were enhanced by 99.07% (P<0.05) and 18.38%(P<0.05), respectively.

关键词： pig rice β-glucanase xylanase cellulase growth digestion

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