Effect of complex nitrogen sources on the production of penicillin acylase byBacillus megaterium

The following complex nitrogen sources were evaluated for the production of penicillin acylase byBacillus megaterium: casein hydrolysate, corn steep liquor, stick water concentrate, blood meal and defatted sunflower meal. Experiments were run in shake flasks at 30‡C and pH 7.0. Best results were obtained with casein hydrolysate: 244 IU/I were produced with a productivity of 20.3 IU/l/h and yield of 717.6 IU/g of nitrogen. The lowest results correspond to sunflower meal with 39 IU/1.     

林可霉素生物合成培养基的优化

以花生粉和棉籽蛋白粉取代了原培养基中的黄豆饼粉,采用响应面法对林可霉素产生菌的发酵培养基进行了优化.首先通过单因素试验及正交实验确定替代氮源及其浓度,采用Plackett-Burman实验分析各因素的主效应,选出对响应值影响较大的3个因素,即花生粉、K2HPO4和玉米浆.对这些因素做爬坡实验,确定三个重要因素的中心点浓...     

Improvement of Omega-3 Docosahexaenoic Acid Production by Marine Dinoflagellate Crypthecodinium cohnii Using Rapeseed Meal Hydrolysate and Waste Molasses as Feedstock

Rapeseed meal and waste molasses are two important agro-industrial by-products which are produced in large quantities. In this study, solid state fermentation and fungal autolysis were performed to produce rapeseed meal hydrolysate (RMH) using fungal strains of Aspergillus oryzae, Penicillium oxalicum and Neurospora crassa. The hydrolysate was used as fermentation feedstock for heterotrophic growth of microalga Crypthecodinium cohnii that produce docosahexaenoic acid (DHA). The addition of waste molasses as a supplementary carbon source greatly increased the biomass and DHA yield. In the batch fermentations using media composed of diluted RMH (7%) and 1-9% waste molasses, the highest biomass concentration and DHA yield reached 3.43 g/L and 8.72 mg/L, respectively. The algal biomass produced from RMH and molasses medium also had a high percentage of DHA (22-34%) in total fatty acids similar to that of commercial algal biomass. RMH was shown to be rich in nitrogen supply comparable to the commercial nitrogen feedstock like yeast extract. Using RMH as sole nitrogen source, waste molasses excelled other carbon sources and produced the highest concentration of biomass. This study suggests that DHA production of the marine dinoflagellate C. cohnii could be greatly improved by concomitantly using the cheap by-products rapeseed meal hydrolysate and molasses as alternative feedstock.     

Induction of collagenase production in Vibrio B-30.

The inducible nature of an extracellular collagenase produced by a marine Vibrio (Vibrio B-30, ATCC 21250) was demonstrated by observing the increase in extracellular collagenase activity after the addition of collagen to cell cultures in the latter part of the exponential growth phase. When collagenase-hydrolyzed collagen was added, the lag time required before collagenase production was detected decreased significantly compared with cultures receiving collagen. Cells preinduced to synthesize collagenase did not produce the enzyme when collagen was removed from the culture medium. Incorporation of penicillin G had no effect on final collagenase activity levels in suspensions of Vibrio B-30 in complete medium supplemented with collagen. However, chloramphenicol and tetracycline inhibited collagenase production, indicating that de novo protein synthesis was necessary for the appearance of activity. Attempts to isolate the inducing substance(s) involved filtering hydrolyzed collagen through a series of ultrafiltration membranes. The lowest-molecular-weight fraction of collagen hydrolysate with inducing ability was between 1,000 and 10,000. Gel filtration of this fraction on Sephadex G-50 resulted in the appearance of three protein peaks, two of which were capable of inducing collagenase production. Results from amino acid composition and N-terminal amino acid analysis suggest that the inducing substance originates from the polar helical portion of the collagen molecule.     

鱼溶浆、酶解鱼溶浆和酶解鱼浆完全替代鱼粉对黄颡鱼生长的影响

为了探究鱼溶浆、酶解鱼溶浆和酶解鱼浆完全替代鱼粉后对黄颡鱼(Pelteobagrus fulvidraco)生长的影响, 在黄颡鱼实用型日粮配方模式下, 以28%的日本级蒸汽鱼粉为对照(FM), 按照FM组鱼粉蛋白质量的15%、30%和45%三个梯度, 分别添加鱼溶浆(SW15、SW30和SW45)、酶解鱼溶浆(SWH15、SWH30和SWH45)、酶解鱼浆(FPH15、FPH30和FPH45)完全替代鱼粉, 设计共10组等氮等能日粮, 每组设定3个重复, 在池塘网箱中养殖初始体重(18.68±0.10) g /尾黄颡鱼60d。结果显示: (1)以FM为对照, SW的3个试验组黄颡鱼的SGR降低9.89%—20.33%(P<0.05), FCR增加18.79%—44.85%(P<0.05); SWH试验组中SWH15和SWH45的SGR相对FM组分别降低8.79%和14.84%, FCR相对FM组增加16.97%和27.88%, 差异显著(P<0.05); FPH试验组中FPH15和FPH45的SGR相对FM组降低13.18%和15.38%, FCR相对FM增加30.30%和29.70%, 差异显著(P<0.05); (2)SWH30和FPH30组的SGR、FCR与FM组无显著差异(P>0.05); (3)相对于SW, 同等添加量SWH的SGR增加4.40%—9.39%, FCR降低9.81%—15.31%(P<0.05); (4)与FM组的FIFO值相比, SW30、SWH30和FPH30的FIFO值仅为0.67、0.61和0.60, 降低67.67%—70.15%(P<0.05)。结果表明: 在黄颡鱼日粮中, 8.5%添加量的酶解鱼溶浆(干物质)、8.2%添加量的酶解鱼浆(干物质)可以完全替代鱼粉(28%), 而相应的FIFO值分别比鱼粉组降低了69.95%和70.15%; 酶解鱼溶浆对黄颡鱼的生长效果优于鱼溶浆。日粮中过高或过低添加量的鱼溶浆、酶解鱼溶浆和酶解鱼浆都会导致黄颡鱼的生长速度和饲料效率下降。     

Simultaneous inclusion of sorghum and cottonseed meal or millet in broiler diets: effects on performance and nutrient digestibility

Two experiments were conducted to investigate the use of sorghum, cottonseed meal and millet in broiler diets and their interaction when they are used simultaneously. In Experiment 1, a corn-soybean meal control diet was compared with eight experimental treatments based on low tannin sorghum (S30, S45 and S60), cottonseed meal (CM15, CM40) or both ingredients included in the same diet (S30/CM40, S45/CM25 and S60CM15). Results showed that BW gain was not affected by the inclusion of sorghum or cottonseed meal. However, feed intake tended to be affected by the cereal type with the highest values with sorghum-based diets. Feed conversion ratio increased (P<0.001) with sorghum-based diets compared with the control diet, whereas a combination of cottonseed meal and sorghum in the same diet did not affect the feed conversion ratio. Significant differences (P<0.001) were observed in apparent ileal digestibility (%) of protein and energy with the cottonseed meal and sorghum/cottonseed meal-based diets having lower protein and energy digestibility compared with corn-based diets. In Experiment 2, a control diet was compared with six diets in which corn was substituted at 60%, 80% or 100% by either sorghum or millet and other three diets with simultaneous inclusion of these two ingredients (S30/M30, S40/M40, S50/M50). Single or combined inclusion of sorghum and millet resulted in similar feed intake and growth performance as the control diet. Apparent ileal digestibility of protein and energy was higher with millet-based diets (P<0.001). Total tract digestibility of protein in sorghum and millet-based diets tended to decrease linearly with the increasing level of substitution. Sorghum-based diets resulted in lower total tract digestibility of fat compared with millet and sorghum/millet-based diets (P<0.001). Higher total tract digestibility of starch were obtained with the control diet and millet-based diets compared with the sorghum-based treatments. Results of the two experiments suggest that broiler growth performance was not affected by the dietary level of sorghum, millet or cottonseed meal. Nutrient digestion can, however, be affected by these feed ingredients.     

高效棉酚降解枯草芽孢杆菌M-4菌株的常压室温等离子体诱变及发酵工艺优化

【背景】棉粕中游离棉酚的存在制约了棉粕作为饲料蛋白源的利用,棉酚的微生物降解问题成为研究热点。本实验室前期发现枯草芽孢杆菌(Bacillus subtilis) M-4菌株具有较强的降解棉酚能力,而且已经应用于棉粕脱毒工业。【目的】进一步提高枯草芽孢杆菌M-4菌株降解棉粕中棉酚的能力,扩大棉粕在养殖业的应用领域。【方法】采用常压室温等离子体诱变(atmospheric and room temperature plasma,ARTP)技术对菌株M-4进行诱变,以液体培养条件下的棉酚降解率为初筛指标,筛选获得正向突变株。以棉粕固体发酵条件下的棉酚降解率为复筛指标,测定初筛获得的正向突变株实际降解棉粕中游离棉酚的能力。以棉酚残存量和棉酚降解率为检测指标,采用单因素试验优化固体条件下棉粕发酵条件,获得突变株发酵棉粕的最适工艺参数。【结果】初筛获得正向突变株19株。复筛得到一株高效的突变株MY-4-17,在棉粕固体发酵条件下其棉酚降解率高达97.15%,比菌株M-4的棉酚降解率提高了2.55%。经过5次传代培养,突变株MY-4-17的遗传稳定性良好。突变株MY-4-17发酵棉粕的最适工艺参数为...     

Apparent digestibilities and growth experiments with tilapia (Oreochromis niloticus) fed soybean meal, cottonseed meal and sunflower seed meal

Apparent digestibility and growth experiments with three oil seed by-products, soybean meal, cottonseed meal, and sunflower seed meal, were conducted using tilapia ( Oreochromis niloticus ) with an initial average body weight of 93 ± 5 g and 64 ± 1 g, respectively. The diets were formulated on an isonitrogeneous and isoenergetic basis. During the experimental period the fish were fed 1% and 1.5% of their metabolic body weight (kg^0.8) daily in the digestibility experiment and the growth experiment, respectively. Water temperature was maintained at 26.5 ± 1°C. Apparent digestibilities of the components and the diets were determined using an indirect method, i.e. with HCl-insoluble ash as an indicator. Faeces collection was carried out using a sedimentation technique.
Results of the apparent digestibilities showed that the crude protein, crude fat, as well as the gross energy digestibilities of soybean meal (93.0%, 94.6%, and 77.2%, respectively), were better than those of cottonseed meal (79.4%, 83.2%, and 57.9%, respectively) and sunflower seed meal (89.8%, 82.9%, and 49.3%, respectively). With the exception of the crude protein digestibility, nutrient and gross energy digestibilities of the cottonseed meal were found to be better than that of sunflower seed meal.
Growth experiment results indicated that soybean meal diet groups had inferior growth performance and feed conversion in comparison to the fishmeal control diet or the other test diets at an inclusion level of 32% of the digestible protein. Fish fed cottonseed meal diet and sunflower seed meal diet showed a relatively better, but not significantly different performance than did the fishmeal control diet. Supplementation diets with lysine and methionine did not enhance fish performance.     

棉籽粕和菜籽粕替代部分鱼粉对黄颡鱼肉质及肌肉发育相关基因表达的影响

文章旨在研究菜籽粕与棉籽粕复合替代饲料鱼粉对黄颡鱼(Pelteobagrus fulvidraco)肌肉品质及肌纤维发育相关基因表达的影响。实验设计了5组等氮等能的饲料,以含鱼粉25%的处理组为对照饲料(FM),以菜籽粕与棉籽粕3﹕2混合后分别替代其中的10%、20%、30%和40%鱼粉,设置了RM10、RM20、RM30和RM40四个试验组。使用初始体质量为(2.38±0.10) g的黄颡鱼幼鱼,分别用上述5种饲料饲喂10周。试验表明,与对照组相比,菜粕棉粕混合替代水平达到20%及以上时黄颡鱼肌肉中的粗蛋白与粗脂肪显著降低,而水分和灰分则无显著差异;肌肉中的羟脯氨酸含量呈现下降的趋势;肌纤维中直径≥50μm的纤维数量随着替代水平达到20%后不断减小,而≤20μm的纤维数量则呈现上升趋势;植物蛋白源的替代也会对黄颡鱼的肌肉质构产生一定的影响。此外,肌肉纤维发育相关基因myod、myog和mrf4的mRNA水平随着替代水平上升而逐渐升高,而myf5与mstn的基因表达在各组间无显著差异。TOR通路相关基因中的akt、tor和s6k1等基因随着替代水平升高而下降, 4e-bp1则随着替代水...     

Gossypol isomers bind specifically to blood plasma proteins and spermatozoa of rainbow trout fed diets containing cottonseed meal

We investigated the role of gossypol isomers binding to blood plasma, seminal plasma and spermatozoa to elucidate gossypol anti-fertility action in the teleost fish, rainbow trout (Oncorhynchus mykiss). Growth and hematological indicators of males were depressed when fish meal protein in diets was completely replaced with cottonseed meal. The cottonseed meal contained equal proportions of (-) (47.8+/-1.6%) and (+) gossypol isomers. Concentrations of spermatozoa were decreased with increasing proportions of gossypol in diets (from 0.22% to 0.95%); however, sperm motility and fertilizing ability were not affected. In contrast to mammals, steroid hormone concentrations were not suppressed in fish given diets with gradual increase of gossypol level. Gossypol concentrations were 100-fold higher in blood plasma than in seminal plasma, confirming a barrier in gossypol transfer between the general circulation and the testis. Spermatozoa accumulated predominantly (+) enantiomer (65-75%) with decreasing proportions as dietary gossypol concentrations increased. Spermatozoa bound most of the gossypol contained in the semen; however, this did not result in impairment of the sperm motility apparatus. Teleost fish sperm rely on ATP stores that accumulate during maturation as a source of energy during activation. In addition, the duration of sperm movement is short in these fish. As such, we hypothesize that the major action of gossypol on mammalian sperm, which is uncoupling of oxidative phosphorylation, does not impair the energy supply required for flagellar beating in fish spermatozoa.     

发酵产丁二酸过程中废弃细胞的循环利用

对厌氧发酵产丁二酸后的废弃细胞进行破壁处理,考察了以细胞水解液作为有机氮源重新用于丁二酸发酵的可行性。比较了超声破碎、盐溶、酶解3种方法破碎细胞获得的水解液作为氮源发酵产丁二酸的效果,结果表明酶解制得的细胞水解液效果最佳。以总氮含量为1.11g/L的酶解液(相当于10g/L酵母膏)作为氮源发酵,丁二酸产量可达42.0g/L,继续增大酶解液用量对耗糖、产酸能力没有显著提高。将细胞酶解液与5g/L酵母膏联用发酵36h后,丁二酸产量达75.5g/L,且丁二酸生产强度为2.10g/(L·h),比使用10g/L酵母膏时提高了66.7%。因此,厌氧发酵产丁二酸结束后的废弃细胞酶解液可以替代原培养基中50%的酵母膏用于发酵。     

Apparent digestibility of crude protein and gross energy in some plant and animal-based feedstuffs by Clarias isheriensis (Siluriformes: Clariidae) (Sydenham 1980)

Apparent digestibility coefficient (ACD) of dry matter, crude protein and gross energy of white fish meal, blood meal, poultry by-product meal, hydrolysed feather meal, shrimp head meal, meat and bone meal, soybean meal, cottonseed meal, peanut meal, yellow maize, corn starch, cassava meal, sorghum, rice bran and cocoa pod meal were determined for Clarias isheriensis (47.5–51.2 g) fed to satiation with diets containing each test feedstuff in a 30:70 mixture with a reference diet, and chromium oxide as an indicator using the dissection technique. ADC_protein values ranged from 50 to 93 % while ADC_energy ranged from 42 to 98 %. Protein and energy content of animal or plant feed-stuffs were equally digestible to C. isheriensis.     

Biosynthesis of penicillin V acylase by Fusarium sp.: effect of culture conditions

Penicillin V acylase was produced, both intracellularly and extracellularly, by Fusarium sp. SKF 235 grown in submerged fermentation. When neopeptone was added to the medium, >95% of the penicillin V acylase was extracellular. In the absence of a complex organic nitrogen source, the fungus produced low levels of totally intracellular penicillin V acylase. MgSO₄ was essential for synthesis of the enzyme, which was induced by phenoxyacetic acid and penicillin V. The maximum yield of penicillin V acylase was 430 IU/g dry cell wt. The optimum pH value and temperature for the penicillin V acylase were 6.5 and 55°C, respectively.     

Value of lucerne juice and grass juice as sources of protein for the growing pig

Six experiments, involving a total of 282 individually-fed pigs, were completed in which the value of lucerne juice, and in one experiment of grass juice, as a source of protein for the growing pig was determined. The juices were products of a green crop fractionation process.The juice was preserved in three ways and was stored for feeding over several weeks. Pigs weighing up to 54 kg received sufficient juice to replace either all or half of the protein provided by the 7% white-fish meal included in an all-meal control diet. From 54 kg to 90 kg liveweight, juice replaced all of the protein supplied by the 3.5% white-fish meal included in the all-meal diet. In carrying out the replacement, all nutrients in the juice other than protein were ignored.When the juice was given according to its true protein content, as determined every 14 days, and was used to replace 3.5% white-fish meal protein throughout the growing period from 20 to 90 kg liveweight, performance and carcass quality were closely comparable to those of control pigs fed on meal only, and the consumption of white-fish meal was reduced by 70%. When the juice replaced 7% white-fish meal protein, the results were poorer than those of the all-meal control pigs.Satisfactory results were also obtained when the juice was given in conjunction with liquid cheese whey (replacing, on a dry matter basis, 30% of the barley meal in the all-meal control diet).Propionic acid (1%) was ineffective as a preservative of the juice. The addition of 1.4 g/l of sodium metabisulphite together with acidification with HCl to pH 3 significantly reduced the rate of loss of true protein during storage of the juice; if, in addition, the juice was heated to 85°C for a few seconds, loss of true protein over a period of many weeks was negligible.     

Effects of supplementation with corn distillers’ dried grains on animal performance,nitrogen balance,and enteric CH4 emissions of young Nellore bulls fed a high-tropical forage diet

The inclusion of corn-dried distillers’ grains (DDG) could be an alternative supplement to increase animal performance, nitrogen efficiency usage (NEU), and decrease enteric methane (CH₄) emissions. Our goal was to determine whether DDG could replace a traditional supplement (cottonseed meal) without affecting animal performance, N balance, and CH₄ emissions. The experiment was conducted during the forage growing season (December to April), with 15 d adaptation, and a 112 d experimental period. The experimental design was completely randomized with four treatments: a mineral supplement (MS), cottonseed meal supplement (CS), 50% replacement of CS by DDG (50DDG), and 100% replacement of CS by DDG (100DDG). Cottonseed meal and DDG were used as protein supplement. A total of 12 paddocks, 3 per treatment, were used to measure forage mass: morphological and chemical composition of forage, forage allowance, and animal performance. Six animals per treatment were used to evaluate DM intake, digestibility, CH₄ emissions, microbial protein production (MCP), and NEU of each treatment. Eighty-one Young Nellore bulls (48 testers, 12 per treatments and 33 adjusters) with initial BW of 255 ± 5 kg (10–12 months old) were supplemented with each supplement type at a level of 0.3% of BW. Pasture management was continuous stocking with a variable stocking rate (put-and-take). Enteric CH₄ was measured using the gas tracer technique. The MCP was quantified using purine derivatives and the NEU mass balance. No differences were found in nutrient intake (P > 0.228). Individual animal performance and gain per area were higher in the treatments with concentrates compared with that of MS; however, there was no difference among treatments CS, 50DDG, and 100DDG. The ADG was 0.83 for MS and 1.08 kg/animal/d when supplemented (P < 0.05). Gain per hectare was 709 kg/ha for MS and 915 kg/ha when supplemented with concentrates (P < 0.05). There was no difference in CH₄ production among treatments that average 180 g/animal/d; however, CH₄ per kg of gain was reduced with CS. The CH₄ conversion factor averaged 5.91%. There was no difference in the synthesis of MCP and NEU. Corn DDG can replace 100% of cottonseed meal as a protein source for supplementation of young Nellore bulls grazing in tropical pastures without affecting animal performance, NEU, MCP, and CH₄ emissions.     

Phytase production byAspergillus ficuum on semisolid substrate

Summary Phytase production byAspergillus ficuum was studied using solid state cultivation on several cereal grains and legume seeds. The microbial phytase was used to hydrolyze the phytate in soybean meal and cotton seed meal. Wheat bran, soybean meal, cottonseed meal and corn meal supported good fungal growth and yielded a high level of phytase when an adequate amount of moisture was present. The level of phytase production on solid substrate was higher than that obtained by submerged liquid fermentation. Higher levels of phosphorus (more than 10 mg Pi/100 g substrate) in the growth medium (static culture) inhibited phytase synthesis, and the degree of phosphorus inhibition was less apparent in semisolid medium than in liquid medium. A static cultivation on semisolid substrate produced a higher level of phytase (2-20-fold) than that obtained by agitated cultivation. The minimal amount of water required for growth and enzyme production on those substrates was about 15%, while the optimum level for phytase production was between 25 and 35% and that for cell growth was above 50%. Optimum pH for phytase production was between 4 and 6.A ficuum grew well on raw (unheated) substrate containing a minimal amount of water and produced as much phytase as on heated substrate. About half of the phytic acid in soybean meal and cottonseed meal was hydrolyzed by treatment withA. ficuum phytase.     

采用玉米秸秆水解糖和玉米浆发酵生产丁二酸

研究了以玉米秸秆水解糖为碳源,不同氮源条件下琥珀酸放线杆菌Actinobacillus succinogenesSF-9的丁二酸发酵产酸能力。结果表明玉米浆可以替代酵母膏作为丁二酸发酵的廉价氮源。厌氧摇瓶丁二酸发酵单因素试验,得到在初糖浓度50 g/L时,玉米浆的较佳用量为20 g/L。在5 L搅拌罐上,考察了不同初始玉米秸秆水解糖浓度对A.succinogenes SF-9发酵生产丁二酸的影响,结果显示高初始秸秆糖浓度对琥珀酸放线杆菌的生长有抑制作用。采用补料分批发酵,发酵60 h丁二酸的产量达到42.7g/L,丁二酸产率82.7%,生产强度0.81 g/(L·h)。丁二酸的产量和生产强度较分批发酵有明显提高。     

豆粕水解液为氮源发酵产脂肪酶的研究

研究了以豆粕水解液作为氮源,假丝酵母Candida sp．99—125发酵生产脂肪酶的过程。分析水解时间对于产酶的影响,对比豆粕水解前后作为氮源发酵时的产酶规律。在30L发酵罐中批次发酵酶活最高可达6000IU／mL,采用豆油反馈流加之后,发酵脂肪酶活力可达8500IU／mL。     

大口黑鲈对六种非粮蛋白源的表观消化率

实验以0.1%三氧化二钇(Y₂O₃)为外源指示剂, 用“70%基础饲料+30%待测饲料原料”的方法配制实验饲料。测定初始体重为(19.28±0.10) g的大口黑鲈(Micropterus salmoides)对荚膜甲基球菌蛋白、乙醇梭菌蛋白、黄粉虫粉、酶解大豆、小球藻和棉籽浓缩蛋白的干物质、蛋白质、脂肪、能量和氨基酸的表观消化率。结果显示, 6种实验原料干物质的表观消化率在37.27%—86.43%(荚膜甲基球菌蛋白>乙醇梭菌蛋白>酶解大豆>黄粉虫粉>小球藻>棉籽浓缩蛋白), 其中荚膜甲基球菌蛋白干物质的表观消化率最高, 乙醇梭菌蛋白次之, 黄粉虫粉、酶解大豆和小球藻之间没有显著性差异, 均显著高于棉籽浓缩蛋白(P<0.05); 蛋白质的表观消化率在79.97%—88.45%(荚膜甲基球菌蛋白>乙醇梭菌蛋白>黄粉虫粉>酶解大豆>小球藻>棉籽浓缩蛋白), 其中荚膜甲基球菌蛋白和乙醇梭菌蛋白均显著高于酶解大豆、小球藻和棉籽浓缩蛋白(P<0.05); 脂肪的表观消化率在51....