欧文氏杆菌变异菌株木聚糖酶纯化及其酶学性质

从现有309份菌种资源中筛选出木聚糖酶高产菌株(Erwinia),发酵液酶活达到408 U.采用超滤和凝胶层析法从发酵液中分离纯化出电泳纯木聚糖酶,回收率高达69.17%,比活达到28453.6U/mg.酶学性质研究结果显示,采用SDS-PAGE和凝胶层析测得分子量分别为22.54和21.89 kD;IEF-PAGE测得等电点为9.63;以燕麦木聚精为底物时,K_m和V_(max)分别为0.5mg/mL和533 U;最适作用pH 5.8,稳定pH 3.4～6.4;最适作用温度60℃,在pH 5.8条件下稳定温度0～65℃;Fe~(2+)和Co~(2+)有激活作用,Cu~(2+)有强烈抑制作用.用ESI-MS/MS法分析,该菌株分泌的木聚糖酶氨基酸序列与GenBank上登录的木聚糖酶基因表达的氨基酸序列一致.     

源于厌氧真菌Orpinomyces sp.PC-2的xynA在毕赤酵母中的高效表达及其酶学性质分析

β-D-1,4-内切木聚糖酶(endo-1,4-β-D-xylanase,xynA,EC.3.2.1.8)简称β-木聚糖酶(β-xylanase),是木聚糖降解酶系中最重要的成员,广泛用于饲料、食品、造纸和生物能源等领域.厌氧真菌Orpinomyces sp.PC-2 xynA具有潜在的开发价值,但是其异源表达的活性较低.本研究根据毕赤酵母(Pichia pastoris)对基因密码子偏好性,对厌氧真菌Orpinomyces sp.PC-2的xynA进行密码子优化.通过全基因合成技术合成优化后的xynAm,并克隆到毕赤酵母表达载体pPIC9K的多克隆位点,构建重组质粒pPIC9K-xynAm,电击转化至毕赤酵母GS115菌株中,以29℃、0.5％甲醇诱导表达,获得重组xynA,并对该酶的酶学特性进行了研究.结果表明,在摇瓶水平条件下,诱导表达108 h时重组β-木聚糖酶活性达到最大值,为612 IU/mL.在10L发酵罐中诱导表达96 h后,xynA的活性为3 515 IU/mL,比活性为2 411 IU/mg;菌体培养湿重、干重和培养上清总蛋白质的浓度分别为324、156和2.25 g/L.非变性十二烷基硫酸钠-聚丙烯酰胺(sodiumdodecyl sulfate-polyacrylamide gel electropheresis,SDS-PAGE)凝胶电泳显示,重组木聚糖酶的分子量约为42.7 kD.酶学性质分析表明,重组β-木聚糖酶的最适反应温度为55℃,最适反应pH 6.0,在温度为30～50℃、pH 4.0～10.6之间该酶具有较好的稳定性,其Km=27.86 mg/mL,Vmax=277.78 mg/(mL· min).底物特异性分析表明,重组xynA可水解低粘度阿拉伯木聚糖、高粘度阿拉伯木聚糖、燕麦木聚糖、桦木木聚糖和4-O-甲基-D-葡萄糖醛酸-D-木聚糖,但不降解大麦(Hordeum vulgare)β-葡聚糖和地衣多糖.10 mmol/L的Co2+和K+对该酶的活性略有激活作用,Mn2+、Fe2+和SDS对该酶活性有一定的抑制作用.本研究为进一步工业化应用来源于厌氧真菌Orpinomyces sp.PC-2的β-内切木聚糖酶奠定了基础.     

欧文氏杆菌木聚糖酶的纯化与酶学性质研究

从实验室现有菌种资源中筛选出木聚糖酶高产菌株,发酵液酶活达到408 U;仅采用超滤和凝胶层析2个步骤即从发酵液中提取到电泳纯木聚糖酶,回收率高达69.17 %,比活达到28453.6 U/mg;用ESI-MS/MS法测得该酶氨基酸序列,与该研究室登陆GenBank的木聚糖酶基因表达的氨基酸序列一致;其酶学性质为：采用SDS-PAGE和凝胶层析测得分子量为22.54 kD、21.89 kD;IEF-PAGE测得等电点为9.63;以桦木木聚糖为底物时,Km值和Vmax分别为1.0mg/ml和33.3μmol/（min.ml）,燕麦木聚糖的Km值和Vmax分别为0.5mg/ml和33.3μmol/（min.ml）;最适作用温度60℃,稳定温度0℃~60℃;最适pH5.8,稳定pH3.4~6.4;Fe2+、Co2+有激活作用,Cu2+有强烈抑制作用。     

重组多功能木聚糖酶酶解条件的优化

重组多功能木聚糖酶(recombinant multi-functional xylanase,RMFXase)是以草菇为表达载体,具有多种酶活力的新型酶。为揭示该酶与底物的作用规律,同时为转基因酶的应用提供基础数据,该文利用重组多功能木聚糖酶水解南方特色的甘蔗加工废弃物生产低聚木糖,通过响应面法建立以重组多功能木聚糖酶质量浓度、底物质量浓度和酶解时间为控制因素的反应动力学方程组,从而推导得到酶解最佳工艺参数。试验并通过高效液相色谱法法,监测反应过程中产物的动态变化,分析产物的生成规律。结果表明,通过响应面法推导出符合连续式酶解反应器的最佳酶解工艺参数为:加酶量900 U/mL、底物质量浓度110 mg/mL、反应时间80 min。该条件下酶解木聚糖含量为54.7%的蔗渣半纤维素,得到的低聚木糖DP值达到3,酶解率达到23.94%。酶解过程中重组多功能木聚糖酶表现出高内切-β-1,4-木聚糖活力,优先降解大分子木聚糖,后期产物主要以木二糖和木三糖为主,且底物纯度越高,底物利用率越高。     

黑曲霉发酵麦麸生产β-葡萄糖苷酶的工艺优化及动力学研究

用单因素和正交试验对黑曲霉M85以小麦麸皮为发酵基质生产β-葡萄糖苷酶的工艺条件进行了优化，并研究了最优发酵条件下的动力学模型。正交试验结果表明，培养基中麦麸的有效添加量为2%，适合摇瓶和5 L罐最佳发酵工艺条件：转速分别为(200±5)r/min和(400±10)r/min，发酵温度均为(30±0.5)℃，接种量分别为15%和10%。黑曲霉M85在摇瓶和5 L罐发酵过程中动力学曲线具有相同的变化趋势，发酵第3 d发酵液中β-葡萄糖苷酶酶活分别达到1103.73 U/mL和1318.82 U/mL，对麦麸的转化率分别为55186.61 U/g和76085.82 U/g。其细胞生长和产物合成可以分别用Monod方程和Luedeking-Piret方程拟合。结果可为麦麸资源生物发酵深加工生产β-葡萄糖苷酶提供技术基础。     

黑曲霉固态发酵苹果渣产木聚糖酶的工艺优化研究

为了获得有较高酶活值的低成本木聚糖酶产品,试验以苹果渣和棉粕为基料研究了黑曲霉(Aspergillus niger SL-05)固态发酵产木聚糖酶的最佳条件.初步试验结果表明:以棉粕和苹果渣(1:1)作为基础氮源和碳源得到的发酵曲的酶活值最高.通过Plackett-Burman试验筛选出了对产酶影响显著的3个因素:尿素、KH2PO4、含水率.进一步试验采用二次回归正交旋转试验设计研究了自变量(尿素、KH2PO4、含水率)对黑曲霉SL-05产酶的影响,通过响应面分析获得了产酶的最佳条件.模型的方差检验显示总回归达到了显著水平(P＜0.05),失拟性检验不显著,说明模型适合并且预测尿素、KH2PO4、含水率对产酶的影响非常有效.在最佳条件(基料棉粕和苹果渣比例为1:1,第2氮源尿素2.6%,无机盐KH2PO4 0.09%,速效碳源葡萄糖2%,含水率62.9%,30℃培养60 h)下,分别获得T5662、30000 U/g的木聚糖酶和纤维素酶的高酶活发酵干曲.以廉价的工农业废料作为基本培养基获得了有较高酶活的产品,经济优势明显.     

温度对麦秸混合物料厌氧干发酵中糖类水解酶活性的影响

水解酶活性直接影响到秸秆厌氧干发酵的沼气产量。本文以麦秸为主要原料, 配制干物质含量(TS)35%的混合发酵物料, 设置20 ℃、25 ℃、30 ℃、35 ℃、40 ℃ 5种不同温度条件, 发酵周期为35 d, 测定发酵过程中料液的纤维素酶、木聚糖酶、淀粉酶和蔗糖酶的活性变化。结果表明, 沼气产气高峰在35 ℃左右, 20~40 ℃时, 随着温度的升高, 发酵后料液的料液干物质含量(TS)、可挥发固体含量(VS)、C/N均明显下降, 发酵产沼气后, 各处理发酵液 pH都降到7.0以下, 其中40 ℃的pH降低幅度最大。纤维素酶活性在发酵初期较低, 峰值出现也较晚, 温度升高明显有利于提高纤维素酶活性。木聚糖酶的起始活性较高, 整个发酵过程中呈先升高后降低并渐趋于平缓的变化趋势, 20~25 ℃处理的木聚糖酶活性在整个发酵过程中没有明显峰值。淀粉酶的起始活性较低, 但随后迅速升高, 30~40 ℃处理在10 d左右达到峰值后开始下降, 此后酶活力的变化幅度较前期平稳。蔗糖酶的起始酶活力较低, 随后升高迅速, 温度在30 ℃以上时酶活性增加更快, 35 ℃时15 d达到峰值后迅速下降, 下降速度明显大于30 ℃和40 ℃处理。由此可见, 温度对秸秆厌氧干发酵过程中不同水解酶活性的影响还是有所差异的。     

杏鲍菇产木聚糖酶固态发酵条件优化及其酶学性质

本文利用啤酒糟为主要原料,以杏鲍菇为菌种进行木聚糖酶发酵研究。通过单因素和正交试验确定最佳培养条件,并对其粗酶的酶学性质进行研究。结果表明,各因素对产酶能力的影响大小依次为:培养时间、料水比、氮源、pH、碳源。固态发酵的最佳培养条件为:碳源为以6∶2∶2配备的啤酒糟、玉米芯、麸皮混合物,氮源为1.5%的酵母膏,pH为5,装瓶量为7g/50ml三角瓶,接种量为25%,培养天数为9d,料水比为1∶1.6。其粗酶液的最适反应温度为50℃,最适pH为6。在pH为4～7时酶的稳定性较好,但木聚糖酶的热稳定性较差,60℃以上时酶活力损失70%以上。     

液氨和过氧化氢预处理对稻草酶解效果的影响机制

稻草是一种重要的木质纤维素资源,可以作为纤维素乙醇转化的原料。该试验通过高温过氧化氢(高温HP)、低温过氧化氢(低温HP)和液氨预处理(liquid ammonia treatment,LAT)3种预处理方式来克服生物质原料的酶解顽抗性,促进稻草酶解转化为可发酵单糖。对预处理后的稻草进行酶解试验,利用高效液相色谱法(highperformanceliquid chromatography,HPLC)定量测定了酶解液中的单糖含量,通过酶解转化率和单糖产量对预处理效果进行了分析比较。试验结果表明高温HP、低温HP和LAT 3种预处理方式均有效提升酶解率,其中LAT预处理的酶解促进作用效果最佳,高温HP预处理次之。稻草在120℃、预处理时间为60 min、30%H2O2水溶液与原料质量比为0.75∶1的高温HP预处理下,在纤维素酶添加量为15U/g时葡聚糖和木聚糖的酶解率分别为61.55%和47.82%,每千克干基稻草原料经144h酶解可生产单糖334.5 g。稻草在90℃、含水率60%、驻留时间为5 min、液氨与原料比例为1∶1的LAT预处理下,在纤维素酶添加量为15 U/g时,葡聚糖和木聚糖的72 h酶解率分别为88.62%和79.29%,每千克干基稻草原料经144 h酶解可生产单糖554.1 g,是未处理原料的2.9倍,总糖回收率达到90%。综上所述,LAT预处理稻草的酶解率显著高于其他单一预处理方法,该研究结果可为稻草制取燃料乙醇提供基础数据。     

脉冲电场促进黑曲霉生长提高产酶活性

为了探讨脉冲电场对黑曲霉生长和糖化酶活性的影响,研究采用响应面法设计脉冲电场工作参数(脉冲强度5～15 kV/cm、脉冲持续时间10～100μs和脉冲数50～99)并对黑曲霉孢子悬液进行处理和培养。结果表明:脉冲强度显著(P0.05)影响菌丝干质量和产糖化酶能力,当脉冲强度为12.975 kV/cm、脉冲宽度为54μs和脉冲数为66,黑曲霉的菌丝干质量和糖化酶活性分别为28.05 mg和18.01 U/mL,比对照(未采用脉冲处理)提高了68.27%和14.71%;酯酶同工酶分析未显示出PEF对黑曲霉有诱变作用,但PEF处理明显刺激了黑曲霉孢子萌发,使孢子萌发率增加了77.8%～231%。该研究为黑曲霉菌等霉菌生物学性能的提高及其在实际生产中的应用提供理论依据。     

Submerged citric acid fermentation on orange peel autohydrolysate

The citrus-processing industry generates in the Mediterranean area huge amounts of orange peel as a byproduct from the industrial extraction of citrus juices. To reduce its environmental impact as well as to provide an extra profit, this residue was investigated in this study as an alternative substrate for the fermentative production of citric acid. Orange peel contained 16.9% soluble sugars, 9.21% cellulose, 10.5% hemicellulose, and 42.5% pectin as the most important components. To get solutions rich in soluble and starchy sugars to be used as a carbon source for citric acid fermentation, this raw material was submitted to autohydrolysis, a process that does not make use of any acidic catalyst. Liquors obtained by this process under optimum conditions (temperature of 130 degrees C and a liquid/solid ratio of 8.0 g/g) contained 38.2 g/L free sugars (8.3 g/L sucrose, 13.7 g/L glucose, and 16.2 g/L fructose) and significant amounts of metals, particularly Mg, Ca, Zn, and K. Without additional nutrients, these liquors were employed for citric acid production by Aspergillus niger CECT 2090 (ATCC 9142, NRRL 599). Addition of calcium carbonate enhanced citric acid production because it prevented progressive acidification of the medium. Moreover, the influence of methanol addition on citric acid formation was investigated. Under the best conditions (40 mL of methanol/kg of medium), an effective conversion of sugars into citric acid was ensured (maximum citric acid concentration of 9.2 g/L, volumetric productivity of 0.128 g/(L.h), and yield of product on consumed sugars of 0.53 g/g), hence demonstrating the potential of orange peel wastes as an alternative raw material for citric acid fermentation.     

发酵柑桔皮渣流化干燥传热传质分析

为了开发高效合理的柑桔皮渣发酵—干燥成套设备,需要进行发酵柑桔皮渣的干燥机理分析、传热传质的研究。该文采用流化干燥方法对发酵柑桔皮渣的干燥进行试验研究,建立了小型的流化床干燥试验台,分析了风速、颗粒粒径、初始含水率等对发酵柑桔皮渣流化干燥过程中传热传质的影响。试验表明：流化床的传热传质性能与流体的物理性质、操作参数、颗粒本身的物理性质以及流化床的特性密切相关。试验结果表明,传热系数沿流化床床高增加而减小,在床高4～6 cm之间,传热系数减小的幅度较大;在风速、颗粒粒径、初始含水率3个影响因素中,风速对传热系数的影响最大,当风速从2.06 m/s增大到2.75 m/s时,床高2～4 cm区域的平均局部传热系数增大了近92%。根据试验结果建立了传热传质数学模型并获得了传热无因次准则方程式,为强化传热传质以提高干燥效果提供了理论依据。     

六种果皮原料果胶的理化及凝胶特性比较

为了解不同品种水果的果皮(柚子皮、西番莲皮、脐橙皮、石榴皮、榴莲皮)以及向日葵盘所提取果胶的理化和质构特性,研究了不同原料果胶的得率、色泽、果胶酸含量、甲氧基含量、酯化度、黏度及质构特性,特别是采用高效液相色谱准确测定了各类果胶的分子量。结果表明:柚子皮、向日葵盘和脐橙皮果胶质量分数较高,分别为18.06%、14.61%和14.43%;西番莲皮果胶质量分数为8.76%;而石榴皮及榴莲皮果胶质量分数较低(均<3%)。从分子量看,石榴皮、脐橙皮果胶分子量较大(>1000kDa),向日葵盘果胶分子量最小(483kDa)。此外,几种果胶的溶胶均属低黏度值果胶(<25厘泊),且在pH值为7.0时黏度最大、在pH值为5.0时黏度最小。结合凝胶质构分析表明:石榴皮果胶分子量最大,凝胶强度最大,但为高甲氧基果胶,且得率较低;而向日葵盘果胶分子量最小,但得率较高,且为低甲基果胶,在非糖及含糖体系中均可形成性能优良的凝胶,因此是生产果胶的良好原料。该文为果胶的生产及应用提供参考。     

Inhibitors of 15-lipoxygenase from orange peel

A series of polymethoxylated flavonoids has been isolated from orange peel, and their inhibitory activity toward soybean 15-lipoxygenase was determined. The strongest inhibition was shown by 3,5,6,7,3',4'-hexamethoxyflavone (IC(50) = 49 +/- 5 microM). Sinensetin, nobiletin, tangeretin, tetramethylscutellarein, and 3,5, 6,7,8,3',4'-heptamethoxyflavone were somewhat less active, with IC(50) values of 70-86 microM, comparable to the positive control quercetin (IC(50) = 68 +/- 5 microM). Demethylation apparently results in less active compounds, with 5-O-demethylsinensetin having an IC(50) value of 144 +/- 10 microM. Some other orange peel constituents were isolated and tested as well, hesperidin (IC(50) = 180 +/- 10 microM) and ferulic acid (111 +/- 2 microM), showing moderate activity. The polymethoxylated flavonoids were virtually inactive as scavengers of the diphenylpicrylhydrazyl radical. Hesperidin was only slightly active (24.2 +/- 0.7% scavenged at a concentration of 2 mM), and ferulic acid showed good activity (IC(50) = 86.4 +/- 0.7 microM). From this, it appears that orange peel constituents may counteract enzymatic lipid peroxidation processes catalyzed by 15-lipoxygenase in vitro. The radical scavenging activity of orange peel extracts is only modest.     

秸秆纤维素降解真菌QSH3-3的筛选及其特性研究

为了获得高效降解秸秆纤维素的微生物菌株,采用滤纸降解法和刚果红染色法从含纤维素类物质的土壤中筛选到一株产纤维素酶菌株QSH3-3,通过形态观察和ITS序列分析,鉴定为草酸青霉Penicillium oxalicum QSH3-3。摇瓶产酶试验结果表明,该菌株的最佳产酶条件为:碳源为0.5%的碱处理过的玉米秸秆粉,氮源为0.2% 硫酸铵,起始pH为7,接种量为5%,产酶温度为30℃,培养时间为4 d。最佳产酶条件下,滤纸酶（FPase）、内切酶（CMCase）和木聚糖酶（Xylanase）分别为12 U、33 U、605 U（U为酶活性单位）;在15℃,其残余酶活力可达70%～80%;在pH 4～9 范围内,其残余酶活力可达70%以上。酶学稳定性研究表明,FPase、CMCase和Xylanase在pH 4～9范围残余酶活力达85%以上,具有较强的酸碱适应能力;FPase、CMCase和Xylanase在45℃以上酶活力迅速下降,耐热性较差。该菌株具有较高的木聚糖酶活力以及较强的低温、pH的耐受力,因而该菌株在田间温差大、土壤偏碱性等复杂条件下对秸秆纤维素类物质的降解具有较高的应用潜力。     

Fermented Wheat Bran as a Functional Ingredient in Baking

The aim of the current study was to identify factors influencing the technological functionality of fermented bran. The influences of fermentation type and type of wheat bran on the microbial community, bioactivity, arabinoxylans (AX), and activity of xylanases were studied in the bran ferments. Furthermore, technological quality of ferments was established by using them to replace wheat in baking with a 20% substitution level. Solubilization of AX and endogenous xylanase activity of bran were influenced by the type of bran, fermentation type, and conditions. Peeled bran had a clearly reduced microbial load and different microbial community in comparison to native bran. Bran from peeled kernels contained 10‐fold lower activities of endogenous xylanases in comparison to native bran. Yeast fermentation of bran from peeled kernels increased the level of folates (+40%), free phenolic acids (+500%), and soluble AX (+60%). Bread containing yeast‐fermented peeled bran had improved volume (+10–15%) and crumb softness (25–35% softer) in comparison to unfermented counterparts. Solubilization of AX during the 20 hr fermentation and decreased endogenous xylanase activity are proposed as the main reasons for the improved technological functionality of fermented bran.     

Solid-state fermentation of soybean and corn processing coproducts for potential feed improvement

Two agro-industrial coproducts, soybean cotyledon fiber and distiller's dried grains with solubles (DDGS), were used as substrates to evaluate the effect of coculturing three different fungi, Aspergillus oryzae , Trichoderma reesei , and Phanerochaete chrysosporium , on enzyme production by solid-state fermentation (SSF). When soybean fiber was used as the substrate, a maximum xylanase activity of 757.4 IU/g and a cellulase activity of 3.2 IU/g were achieved with the inoculation and incubation of T. reesei and P. chrysosporium for 36 h, followed by A. oryzae for an additional 108 h. This inoculation scheme also resulted in the highest xylanase activity of 399.2 IU/g compared to other fungi combinations in the SSF of DDGS. A large-scale SSF by this fungus combination produced fermented products that had xylanase and cellulase activities of 35.9-57.0 and 0.4-1.2 IU/g, respectively. These products also had 3.5-15.1% lower fiber and 1.3-4.2% higher protein contents, suggesting a potential feed quality improvement.     

柑桔皱皮果与果皮及其细胞壁矿质元素关系的研究

对红 1 7甜橙、红江橙及红江橙正常果与皱皮果、红江橙阴面果与阳面果的果皮及其细胞壁N、P、K、Ca、Mg的含量进行了分析 ,探讨这些矿质元素与柑桔成熟期皱皮果形成的关系。结果发现红 1-7甜橙皱皮果发生率较低 ,仅为 7.34% ,而红江橙皱皮果发生率高达 28.06% ;皱皮现象 86%以上出现在果实的阴面果皮。果皮N、P含量与柑桔皱皮果的形成没有相关性 ,K在皱皮果中的含量比正常果高 ,而在阳面果皮比阴面果皮高 ;Mg在皱皮果和阴面果皮及其细胞壁的含量都比在正常果和阳面果皮稍低 ,但Mg的含量与皱皮果率的相关性不显著 ;Ca在正常果和阳面果皮及其细胞壁的含量都明显高于皱皮果和阴面果皮 ,皱皮果率与Ca的含量呈显著的负相关     

Quality and elemental content of citrus fruit from exposed and internal canopy positions 1

Quality and elemental content of fruit from internal tree canopies were compared with those from external canopy positions in 4 citrus varieties: ‘Kinnow’ mandarin; ‘Redblush’ grapefruit; ‘Valencia’ orange; and ‘Lisbon’ lemon. Fruit weight, total juice per fruit, peel fresh and dry weight, and rind thickness of fruit from internal canopies of all 4 varieties were significantly higher compared with external fruit. Mandarin, grapefruit, and orange fruit from external canopies had higher soluble solids and specific gravity. Fruit from internal canopies of all varieties had generally higher peel concentrations (% dry weight) of N, P and K due to a dilution effect, while the opposite condition existed in mandarin when these elements were expressed on a percent fresh weight basis. Peel Mg and S from external fruit were higher in all varieties, expressed as percentages of either dry weight or fresh weight. Nitrogen content of mandarin and orange juice and Ca content of grapefruit and lemon juice from external fruit were significantly higher compared to those from internal canopy fruit. Elimination of fruit quality and mineral variations as a result of canopy positions is recommended by the means of cultural practices.     

Fruit quality and mineral nutrient from exposed verses internal canopy positions of four scitrus varieties

>Quality and mineral content of fruit from internal tree canopies were compared with those from external canopy positions in 4 citrus varities: ‘Kinnow’ mandarin; ‘Redblush’ grapefruit; ‘Valencia’ orange; and ‘Lisbon’ lemon. Fruit weight, total juice per fruit, peel fresh and dry weight, and rind thickness of fruit from internal canopies of all 4 varieties were significantly higher compared with external fruit. Mandarin, grapefruit, and orange fruit from external canopies had higher soluble solids and specific gravity. Fruit from internal canopies of all varieties had generally higher peel concentrations (% dry weight) of N, P and K due to a dilution effect, while the opposite condition existed in mandarin when these elements were expressed on a percent fresh weight basis. Peel Mg and S from external fruit were higher in all varieties, expressed as percentages of either dry weight or fresh weight. Nitrogen content of mandarin and orange juice and Ca content of grapefruit and lemon juice from external fruit were significantly higher compared to those from internal canopy fruit. Elimination of fruit quality and mineral variations as a result of canopy positions is recommended by the means of cultural practices.