HPLC—ELSD测定饮料中果糖、葡萄糖、蔗糖和乳糖的含量

为建立一种简便、准确、灵敏测定饮料中果糖、葡萄糖、蔗糖和乳糖含量的高效液相色谱蒸发光散射检测方法。使用Phenomenex-NH2(250 mm×4.6 mm,5 μm)色谱柱分离,以乙腈∶水(80∶20,V∶V)为流动相,流速为1.5 mL/min,ELSD为检测器,漂移管温度45℃,N2流量3.5 bar。结果表明：果糖、葡萄糖、蔗糖、乳糖在50～2 000 μg/mL内呈良好线性关系,4种糖的回收率均为96.1%～100.4%,测定的相对标准偏差为0.2%～0.9%。此方法操作简便、快捷、准确,用于实际样品的测定后,获得了令人满意的结果。     

高产纳豆激酶突变菌株产酶条件的研究

为了获得最佳产酶条件,通过研究不同碳源(葡萄糖、木糖、麦芽糖、蔗糖、淀粉),氮源(牛肉膏、蛋白胨、大豆蛋白胨、酵母膏、硫酸铵),碳氮浓度比和无机离子组成的培养基发酵,利用纤维蛋白平板法测定纳豆激酶的活性.得到纳豆激酶的最佳产酶条件为:1.5%蛋白胨,1.5%麦芽糖,0.05%硫酸镁,0.2%氯化钙,0.2%磷酸二氢钠和0.1%磷酸氢二钠;碳氮浓度比为1:1.     

添加甘蔗糖蜜对湖泊红球藻及其虾青素合成的影响

虾青素(Astaxanthin)是类胡萝卜素的含氧衍生物,具有较强的抗氧化、清除自由基等功能;湖泊红球藻(Haematococcus lacustris)是天然虾青素的最好来源之一.为了明确用甘蔗糖蜜或葡萄糖为培养基碳源对湖泊红球藻生长及其合成虾青素的影响,在BG11培养基中分别添加浓度为0.5、1、1.5 g/L的糖...     

解淀粉芽孢杆菌TWC2发酵条件的优化

为了提高解淀粉芽孢杆菌Bacillus amyloliquefaciens TWC2菌株活性代谢物质的产量,以菌体生物量和发酵液拮抗甘蔗赤腐病菌(Colletotrichum falcatum Went.)活性为指标,采用单因素和正交试验的方法对TWC2菌株发酵培养基成分和发酵条件进行优化。结果表明,TWC2菌株最佳发酵培养基组成为:5 g/L麦芽糖,10 g/L果糖,10 g/L蛋白胨,10 g/L酵母浸出粉,1 g/L CaCl_2。最适发酵条件为:培养温度34℃,摇床转速200 r/min,发酵起始pH 6.0~6.5,接种量7%,装液量40 mL/250 mL,发酵时间为48 h。在最佳发酵培养基和培养条件下,发酵液抑制甘蔗赤腐病菌的能力较优化前提高了39.91%。     

金龟子绿僵菌菌株培养基的改良

采用液体振荡与固体平板双培养的方法,对金龟子绿僵菌MA4,MAIml两菌株的培养基进行改良.研究结果表明:MA4,MAIml菌株的最适复合碳源均为玉米粉、最适氮源为蛋白胨、最适碳氮比为4:1(m:m,下同)、碳浓度分别为0.5,1.0mol/L,即改PPDA培养基为:MA4菌株以11.8g/L的玉米粉代替蔗糖,17.1g/L的酵母浸膏代替蛋白胨;而MAlml菌株的则以23.6g/L的玉米粉代替蔗糖,蛋白胨由10.0g/L更换为16.2g/L.     

巴西橡胶树红根病病菌液体培养条件优化研究

通过设置不同碳源、氮源、温度、菌龄等对巴西橡胶树红根病病菌[Ganoderma pseudoferreum (Wakef) Over.et Steinm]进行液体培养试验。结果表明：适合橡胶树红根病病菌菌丝生长的液体培养基配方为：蔗糖1%,葡萄糖2%,D-果糖2%,玉米粉0.5%,酵母粉0.5%,黄豆粉0.8%,蛋白胨1%,KH2PO4 0.1%,MgSO4 0.1%,VB1 0.02%;最适培养温度是29℃,液体菌种菌龄为120 h,培养周期为240 h。     

人参内生菌B69菌株抑菌活性物质产生条件的优化

目的探索人参内生菌B69菌株对根腐病的抑制作用,优化人参病害拮抗菌株B69的发酵条件,提高发酵液的活菌含量和抗菌活性。方法以B69发酵液对病原菌根腐病的抑制作用为活性指标,采用正交试验和单因素试验方法对菌株的最适发酵培养基成分及发酵条件进行优化。结果在最佳发酵培养基和培养条件下,经检测发酵液的活菌数含量为8.25×108 cfu/m L,较优化前的活菌含量1.82×108cfu/m L增幅达到了显著差异(P0.05),此含量达到了农业部微生物肥料的技术标准(有效活菌数≥2.0亿/m L)。发酵液对人参根腐病的抑菌圈也由优化前的18mm增加到35mm。结论最佳培养基配方为蔗糖2.0%、淀粉1.0%、牛肉浸膏0.5%、酵母粉0.5%、蛋白胨0.2%和Na Cl0.5%;初始p H7.0;最适装液量为100 m L/250 m L;培养条件为35℃、170r/min振荡培养;接种量5%,发酵时间为48h。     

米曲霉40214高产中性蛋白酶发酵条件的优化

运用现代生物学技术,对米曲霉40214菌株发酵产中性蛋白酶的培养基组分及培养条件进行优化,旨在提高菌株产中性蛋白酶的产量。通过单因素实验和正交实验确定了发酵培养基的最佳组分,添加葡萄糖8.0%,酵母粉4.0%,Na2HPO40.10%;最适发酵条件为发酵温度28℃,初始pH值6.5,发酵时间96 h,加水量60.0%,经过优化后酶活性提高了1.75倍,达到2 845.68 U/mL,实验为进一步提高米曲霉40214菌株产中性蛋白酶的能力并将其应用到食品发酵中奠定了理论基础。     

粤西1号芒果成熟阶段的蔗糖代谢

以粤西1号为试材研究了芒果果实成熟阶段淀粉、葡萄糖、果糖和蔗糖的含量以及蔗糖代谢相关酶-酸性转化酶(AI)、中性转化酶(NI)、蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)的活性变化.结果表明:采收前淀粉含量逐渐减少,采收后快速减少;葡萄糖含量采收后呈先增后减的趋势,果糖含量采收前缓慢增加,采收后开始迅速增加,蔗糖变化与果糖变化趋势一致,完熟时含量最多的糖为蔗糖(62.55 mg,g),果糖次之(44.45 mg/g),葡萄糖最少(2.87mg/g).AI、NI、SS、SPS在采收前活性差别不大,在后熟阶段,AI、SS、SPS活性显著增加,且SPS活性达到最高值.粤西1号糖代谢以积累蔗糖为主,2种酶SPS和SS起关键作用.     

特高糖抗寒甘蔗新品种桂糖28的选育

为培育高产、高糖、多抗的甘蔗新品种,不断满足糖料生产发展的需要,2000～2008年,采用"五圃制"的常规杂交育种的技术路线进行选择与培育,育成了特高糖高产抗旱抗寒的甘蔗新品种桂糖28。桂糖28(原名桂糖00-122),其亲系为CP80-1018×CP88-2032。该品种中至中大茎,特早熟、特高糖、高产、宿根性好,抗逆性强,适应性广,极易脱叶。甘蔗蔗糖分(16.60%)较ROC16(15.07%)高1.53%,最高甘蔗蔗糖分达19.19%;产蔗量(86.2 t/hm2)较ROC16(83.1 t/hm2)增产3.73%,最高产蔗量达159.2 t/hm2;含糖量(14.31 t/hm2)较ROC16(12.52 t/hm2)增产14.30%。适应于广西、云南、广东、福建等省蔗区种植。桂糖28的育成是广西甘蔗高糖育种取得的突破性进展,对丰富我国甘蔗品种材料,改良目前甘蔗单一化的品种结构具有重要现实意义。     

Development in Aspergillus

The genus Aspergillus represents a diverse group of fungi that are among the most abundant fungi in the world. Germination of a spore can lead to a vegetative mycelium that colonizes a substrate. The hyphae within the mycelium are highly heterogeneous with respect to gene expression, growth, and secretion. Aspergilli can reproduce both asexually and sexually. To this end, conidiophores and ascocarps are produced that form conidia and ascospores, respectively. This review describes the molecular mechanisms underlying growth and development of Aspergillus.Key words: Aspergillus, fungi, asexual reproduction, sexual reproduction, development, conidium, conidiophore, vegetative mycelium, heterogeneity, ascocarp, ascospore, fruiting body     

黑曲霉菌渣资源化再利用途径分析

黑曲霉菌渣是黑曲霉发酵法生产葡萄糖酸钠后产生的废弃物，其资源化利用已成为企业扩大生产亟需解决的问题。对该菌渣有效成分检测得知，该菌渣有害金属元素均小于城镇垃圾农用控制标准、有机-无机复混肥料控制标准和鱼粉、猪、家禽添加剂预混合饲料卫生指标，有机质含量符合有机肥料中的有机质含量指标，总糖、粗纤维、粗蛋白、脂肪等营养成分含量较多，同时也含有丰富的N、P、K、Ca、Mg、Fe等元素，可从有机肥料和禽畜饲料方向进行资源化利用，有利于减轻环境污染及实现变废为宝，具有积极的社会、经济和生态效益。     

New and revisited species in Aspergillus section Nigri

Four new species, Aspergillus eucalypticola, A. neoniger, A. fijiensis and A. indologenus are described and illustrated. Aspergillus eucalypticola was isolated from Eucalyptus leaf from Australia, and is related to A. tubingensis and A. costaricaensis, but could clearly be distinguished from them based on either β-tubulin or calmodulin sequence data. Aspergillus eucalypticola produced pyranonigrin A, funalenone, aurasperone B and other naphtho-γ-pyrones. Aspergillus neoniger is also a biseriate species isolated from desert sand in Namibia, and mangrove water in Venezuela, which produces aurasperone B and pyranonigrin A. Aspergillus fijiensis is a uniseriate species related to A. aculeatinus, and was isolated from soil in Fiji, and from Lactuca sativa in Indonesia. This species is able to grow at 37 °C, and produces asperparalines and okaramins. Aspergillus indologenus was isolated from soil, India. This species also belongs to the uniseriate group of black aspergilli, and was found to be related to, but clearly distinguishable from A. uvarum based on β-tubulin, calmodulin and ITS sequence data. Aspergillus indologenus produced the insecticidal compounds okaramins A, B, H, and two types of indol-alkaloids which have not been structure elucidated. Two other species, A. violaceofuscus and A. acidus, are revalidated based on molecular and extrolite data. Aspergillus violaceofuscus was found to be related to A. japonicus, and produced some of the same interesting indol-alkaloids as A. indologenus, and also produced several families of partially characterised extrolites that were also found in A. heteromorphus. Aspergillus acidus (previously known as A. foetidus var. pallidus and A. foetidus var. acidus) is also a valid species, while A. foetidus is a synonym of A. niger based on molecular and physiological data. Two other species described previously, A. coreanus and A. lacticoffeatus, were found to be colour mutants of A. acidus and A. niger, respectively. Methods which could be used to distinguish the two closely related and economically important species A. niger and A. awamori are also detailed. Although these species differ in their occurrence and several physiological means (elastase activities, abilities to utilise 2-deoxy-D-glucose as sole carbon source), our data indicate that only molecular approaches including sequence analysis of calmodulin or β-tubulin genes, AFLP analysis, UP-PCR analysis or mtDNA RFLP analysis can be used reliably to distinguish these sibling species. Aspergillus section Nigri now includes 26 taxa.     

黄曲霉对花生和玉米的侵染研究

黄曲霉(Aspergillus flavus)极易侵染花生、玉米等多种农作物,并且在侵染后产生具有致癌、致畸、致突变作用的有毒代谢产物——黄曲霉毒素.黄曲霉的侵染和黄曲霉毒素污染不仅发生在作物生长环节,而且在收获、干燥、储运等过程也会发生,严重威胁农产品消费安全和人畜生命健康.因此,解析黄曲霉对粮油作物的侵染过程及侵染...     

New taxa in Aspergillus section Usti

Based on phylogenetic analysis of sequence data, Aspergillus section Usti includes 21 species, inclucing two teleomorphic species Aspergillus heterothallicus (= Emericella heterothallica) and Fennellia monodii. Aspergillus germanicus sp. nov. was isolated from indoor air in Germany. This species has identical ITS sequences with A. insuetusCBS 119.27, but is clearly distinct from that species based on β-tubulin and calmodulin sequence data. This species is unable to grow at 37 °C, similarly to A. keveii and A. insuetus. Aspergillus carlsbadensis sp. nov. was isolated from the Carlsbad Caverns National Park in New Mexico. This taxon is related to, but distinct from a clade including A. calidoustus, A. pseudodeflectus, A. insuetus and A. keveii on all trees. This species is also unable to grow at 37 °C, and acid production was not observed on CREA. Aspergillus californicus sp. nov. is proposed for an isolate from chamise chaparral (Adenostoma fasciculatum) in California. It is related to a clade including A. subsessilis and A. kassunensis on all trees. This species grew well at 37 °C, and acid production was not observed on CREA. The strain CBS 504.65 from soil in Turkey showed to be clearly distinct from the A. deflectus ex-type strain, indicating that this isolate represents a distinct species in this section. We propose the name A. turkensis sp. nov. for this taxon. This species grew, although rather restrictedly at 37 °C, and acid production was not observed on CREA. Isolates from stored maize, South Africa, as a culture contaminant of Bipolaris sorokiniana from indoor air in Finland proved to be related to, but different from A. ustus and A. puniceus. The taxon is proposed as the new species A. pseudoustus. Although supported only by low bootstrap values, F. monodii was found to belong to section Usti based on phylogenetic analysis of either loci BLAST searches to the GenBank database also resulted in closest hits from section Usti. This species obviously does not belong to the Fennellia genus, instead it is a member of the Emericella genus. However, in accordance with the guidelines of the Amsterdam Declaration on fungal nomenclature (Hawksworth et al. 2011), and based on phylogenetic and physiological evidence, we propose the new combination Aspergillus monodii comb. nov. for this taxon. Species assigned to section Usti can be assigned to three chemical groups based on the extrolites. Aspergillus ustus, A. granulosus and A. puniceus produced ustic acid, while A. ustus and A. puniceus also produced austocystins and versicolorins. In the second chemical group, A. pseudodeflectus produced drimans in common with the other species in this group, and also several unique unknown compounds. Aspergillus calidoustus isolates produced drimans and ophiobolins in common with A. insuetus and A. keveii, but also produced austins. Aspergillus insuetus isolates also produced pergillin while A. keveii isolates produced nidulol. In the third chemical group, E. heterothallica has been reported to produce emethallicins, 5'-hydroxyaveranthin, emeheterone, emesterones, 5'-hydroxyaveranthin.     

New species in Aspergillus section Terrei

Section Terrei of Aspergillus was studied using a polyphasic approach including sequence analysis of parts of the β-tubulin and calmodulin genes and the ITS region, macro- and micromorphological analyses and examination of extrolite profiles to describe three new species in this section. Based on phylogenetic analysis of calmodulin and β-tubulin sequences seven lineages were observed among isolates that have previously been treated as A. terreus and its subspecies by Raper & Fennell (1965) and others. Aspergillus alabamensis, A. terreus var. floccosus, A. terreus var. africanus, A. terreus var. aureus, A. hortai and A. terreus NRRL 4017 all represent distinct lineages from the A. terreus clade. Among them, A. terreus var. floccosus, A. terreus NRRL 4017 and A. terreus var. aureus could also be distinguished from A. terreus by using ITS sequence data. New names are proposed for A. terreus var. floccosus, A. terreus var. africanus, A. terreus var. aureus, while Aspergillus hortai is recognised at species level. Aspergillus terreus NRRL 4017 is described as the new species A. pseudoterreus. Also included in section Terrei are some species formerly placed in sections Flavipedes and Versicolores. A. clade including the type isolate of A. niveus (CBS 115.27) constitutes a lineage closely related to A. carneus. Fennellia nivea, the hypothesized teleomorph is not related to this clade. Aspergillus allahabadii, A. niveus var. indicus, and two species originally placed in section Versicolores, A. ambiguus and A. microcysticus, also form well-defined lineages on all trees. Species in Aspergillus section Terrei are producers of a diverse array of secondary metabolites. However, many of the species in the section produce different combinations of the following metabolites: acetylaranotin, asperphenamate, aspochalamins, aspulvinones, asteltoxin, asterric acid, asterriquinones, aszonalenins, atrovenetins, butyrolactones, citreoisocoumarins, citreoviridins, citrinins, decaturins, fulvic acid, geodins, gregatins, mevinolins, serantrypinone, terreic acid (only the precursor 3,6-dihydroxytoluquinone found), terreins, terrequinones, terretonins and territrems. The cholesterol-lowering agent mevinolin was found in A. terreus and A. neoafricanus only. The hepatotoxic extrolite citrinin was found in eight species: A. alabamensis, A. allahabadii, A. carneus, A. floccosus, A. hortai, A. neoindicus, A. niveus and A. pseudoterreus. The neurotoxic extrolite citreoviridin was found in five species: A. neoafricanus, A. aureoterreus, A. pseudoterreus, A. terreus and A. neoniveus. Territrems, tremorgenic extrolites, were found in some strains of A. alabamensis and A. terreus.     

Sesquiterpenoids from the Mangrove-Derived Aspergillus ustus 094102

Four new drimane sesquiterpenoids (1–4) and three known ones (5–7) were isolated from the fermentation broth of the mangrove-derived Aspergillus ustus 094102. Compound 5 was further resolved as four purified compounds 5a–5d. By means of extensive spectroscopic and ECD analysis as well as the chemical transformation, their structures were identified as (2R,3R,5S,9R,10S)-2,3,9,11-tetrahydroxydrim-7-en-6-one (ustusol F, 1), (2R,3R,5R,9S,10R)-2,3,11-trihydroxydrim-7-en-6-one (9-deoxyustusol F, 2), (3S,5R,9R,10R)-3,11,12-trihydroxydrim-7-en-6-one (ustusol G, 3), (5S,6R,9S,10S, 11R,2′E,4′E)-(11-dideoxy-11-hydroxystrobilactone A-6-yl)-5-carboxypenta-2,4-dienoate (ustusolate H, 4), ((5S,6R,9S,10S)-strobilactone A-6-yl) (2E,4E)-6,7-dihydroxyocta-2,4-dienoate (ustusolate I, 5), (2′E,4′E;6′,7′-erythro)-ustusolate I (5a) and (2′E,4′E;ent-6′,7′-erythro)-ustusolate I (5b), (2′E,4′E,6′R,7′R)-ustusolate I (5c) and (2′E,4′E,6′S,7′S)-ustusolate I (5d), (5S,6R,9S,10S,2′E,4′E)-(strobilactone A-6-yl)-5-carboxypenta-2,4-dienoate (ustusolate J, 6), and (2S,5S,9R,10S)-2,9,11-trihydroxydrim-7-en-6-one (ustusol B, 7), respectively. Compound 5 showed antiproliferation against the human tumor cells CAL-62 and MG-63 with the IC₅₀ values of 16.3 and 10.1 µM, respectively.     

Cytosolic streaming in vegetative mycelium and aerial structures of Aspergillus niger

Aspergillus niger forms aerial hyphae and conidiophores after a period of vegetative growth. The hyphae within the mycelium of A. niger are divided by septa. The central pore in these septa allows for cytoplasmic streaming. Here, we studied inter- and intra-compartmental streaming of the reporter protein GFP in A. niger. Expression of the gene encoding nuclear targeted GFP from the gpdA or glaA promoter resulted in strong fluorescence of nuclei within the vegetative hyphae and weak fluorescence in nuclei within the aerial structures. These data and nuclear run on experiments showed that gpdA and glaA are higher expressed in the vegetative mycelium when compared to aerial hyphae, conidiophores and conidia. Notably, gpdA or glaA driven expression of the gene encoding cytosolic GFP resulted in strongly fluorescent vegetative hyphae and aerial structures. Apparently, GFP streams from vegetative hyphae into aerial structures. This was confirmed by monitoring fluorescence of photo-activatable GFP (PA-GFP). In contrast, PA-GFP did not stream from aerial structures to vegetative hyphae. Streaming of PA-GFP within vegetative hyphae or within aerial structures of A. niger occurred at a rate of 10–15 μm s^-1. Taken together, these results not only show that GFP streams from the vegetative mycelium to aerial structures but it also indicates that its encoding RNA is not streaming. Absence of RNA streaming would explain why distinct RNA profiles were found in aerial structures and the vegetative mycelium by nuclear run on analysis and micro-array analysis.Key words: aerial hypha, Aspergillus, conidia, conidiophore, cytoplasmic streaming, development, fungus, vegetative mycelium     

Neuronal Modulators from the Coral-Associated Fungi Aspergillus candidus

Three new p-terphenyl derivatives, named 4″-O-methyl-prenylterphenyllin B (1) and phenylcandilide A and B (17 and 18), and three new indole-diterpene alkaloids, asperindoles E–G (22-24), were isolated together with eighteen known analogues from the fungi Aspergillus candidus associated with the South China Sea gorgonian Junceela fragillis. The structures and absolute configurations of the new compounds were elucidated on the basis of spectroscopic analysis, and DFT/NMR and TDDFT/ECD calculations. In a primary cultured cortical neuronal network, the compounds 6, 9, 14, 17, 18 and 24 modulated spontaneous Ca²⁺ oscillations and 4-aminopyridine hyperexcited neuronal activity. A preliminary structure–activity relationship was discussed.     

Antiviral Cyclopropane Acids from Deep-Sea-Derived Fungus Aspergillus sydowii

Four novel monocyclic cyclopropane acids, namely, sydocyclopropanes A–D (1–4), along with one known congener hamavellone B (5), were isolated from the Aspergillus sydowii MCCC 3A00324 fungus, which was isolated from the deep-sea sediment. The gross structures of novel compounds were established by detailed analyses of the spectroscopic data (HRESIMS and NMR spectra), and their absolute configurations were resolved on the basis of the quantum chemical calculations of ECD and NMR data, in association with DP4+ probability analyses. Sydocyclopropanes A–D, featuring the 1,1,2,3-tetrasubstituted cyclopropane nucleus with different lengthy alkyl side chains, were discovered in nature for the first time. All compounds exhibited antiviral activities against A/WSN/33 (H1N1), with IC₅₀ values ranging from 26.7 to 77.2 μM, of which compound 1 exhibited a moderate inhibitory effect (IC₅₀ = 26.7 μM).