一中国白酒酒曲中的丝状真菌菌群

采用6种不同的培养基,分别于25℃、32℃、42℃以及50℃培养分离从而对某一中国白酒酒曲中的丝状真菌菌群进行研究.从酒曲中共分离得到886株丝状真菌,分属于接合菌,子囊菌和无性型真菌的20属,45种.其中最为丰富的是无性型真菌(28种),其次是接合菌(10种)和子囊菌(7种).对发酵起主要作用的为那些嗜热和耐热的种属,包括:宛氏拟青霉,伞枝梨头霉,梳棉状嗜热丝孢菌,微小根毛霉,金孢霉属一种和红曲属的几个种.文中还对一些在发酵中起重要作用的丝状真菌的特性进行了探讨.     

疏绵状嗜热丝孢菌原生质体的制备与再生

以疏绵状嗜热丝孢菌(Thermomyces lanuginosus)为供试菌株,研究了菌龄、酶的种类及浓度、酶解时间、酶解温度和稳渗剂对原生质体制备的影响及稳渗剂对原生质体再生的影响。结果表明,制备嗜热丝孢菌原生质体比较适宜的条件为:PDB液体培养基培养28 h,以0.7 mol/L NaCl为稳渗剂,0.15 mol/L的溶壁酶,30℃酶解4 h。原生质体再生以0.7 mol/L蔗糖作稳渗剂为最佳。     

Purification and characterization of an extracellular polygalacturonase from the thermophilic fungus,Thermomyces lanuginosus

A polygalacturonase was purified from the thermophilic fungus, Thermomyces lanuginosus to apparent homogeneity by ultrafiltration, acetone precipitation and ion-exchange chromatography. The enzyme was maximally active at pH 5.5 and 60 °C. The apparent K_M with potassium pectate was 0.67 mg/ml and the V_max was 7.2 × 10⁵ mol/min/mg protein. The apparent molecular weight of the enzyme was 59 kDa and it contained approximately 10% carbohydrate. The enzyme was completely stable at room temperature (32 ± 3 °C) and retained about 50% activity at 50 °C for 6 h. The zymogram of the purified enzyme revealed two activity bands, one of which was a major one. Polyclonal antibodies raised against the enzyme did not show any immunological relatedness with other mesophilic polygalacturonases.     

Amylase hyperproduction by deregulated mutants of the thermophilic fungus Thermomyces lanuginosus

Thermomyces lanuginosus was subjected to three cycles of mutagenesis (UV/NTG) and a selection procedure to develop amylase-hyperproducing, catabolite-repression-resistant and partially constitutive strains. One of the selected derepressed mutant strain III₅₁, produced ∼7- and 3-fold higher specific activity of α-amylase (190 U/mg protein) and glucoamylase (105 U/mg protein), respectively, compared to a wild-type parental strain. Further, the effect of production parameters on mutant strain III₅₁ was studied using a Box–Behnken design. The regression models computed showed significantly high R ² values of 96 and 97% for α-amylase and glucoamylase activities, respectively, indicating that they are appropriate for predicting relationships between corn flour, soybean meal and pH with α-amylase and glucoamylase production. Journal of Industrial Microbiology & Biotechnology (2002) 29, 70–74 doi:10.1038/sj.jim.7000270 Received 05 July 2001/ Accepted in revised form 16 April 2002     

Characterization of a xylanase from the newly isolated thermophilic Thermomyces lanuginosus CAU44 and its application in bread making

AIMS: A xylanase from the newly isolated thermophilic fungus, Thermomyces lanuginosus CAU44, was characterized and evaluated for its suitability in bread making. METHODS AND RESULTS: Xylanase was purified 3.5-fold to homogeneity with a recovery yield of 32.8%. It appeared as a single protein band on SDS-PAGE gel with a molecular mass of c. 25.6 kDa. The purified xylanase had an optimum pH of 6.2, and it was stable over pH 5.6-10.3. The optimal temperature of xylanase was 75 degrees C and it was stable up to 65 degrees C at pH 6.2. Study was further carried out to investigate the effect of the purified xylanase on the properties of wheat bread and its staling during storage. CONCLUSIONS: The purified xylanase from T. lanuginosus CAU44 was stable up to 65 degrees C and had a broad pH range. The presence of thermostable xylanase during bread making led to an improvement of the specific bread volume and better crumb texture. Besides, addition of xylanase provided an anti-staling effect. SIGNIFICANCE AND IMPACT OF THE STUDY: The xylanase from the newly isolated Thermomyces lanuginosus CAU44 shows great promise as a processing aid in the bread-making industry.     

利用农业废弃物生产嗜热真菌（T．1anuginosus）耐热木聚糖酶的固体发酵研究

研究了嗜热真菌(Thermomyces lanuginosus CBS288．54-M18)生产木聚糖酶的碳氮源组成、料水比、培养基初始pH值、发酵温度及接种量等的影响。结果表明,以麦麸和玉米芯粉(8：2)作为复合碳源,酵母膏和胰蛋白胨(1：1)作为复合氮源时,菌株所产的木聚糖酶量相比未优化碳氮源的培养基提高幅度高达200％。其最佳产酶的料水比为1：3,培养基初始pH7．0为最适。菌株在50℃条件下发酵5d,能够得到活力高达15023U／g干基碳源的木聚糖酶制剂,且该酶制剂不合纤维素酶和蛋白酶活性。     

Characterization and biobleaching effect of hemicellulases produced by thermophilic fungi

The culture supernatant of Thermomyces lanuginosus strains MED 2D and MED 4B1 had high activities of xylanase with low inducible activities of -xylosidase. The crude xylanase was optimally active at 70 °C and at pH 6.0 to 6.5. Subsequently Eucalyptus kraft pulp was treated with MED 2D supernatant at 10 IU per gram pulp resulting in a 10.5% reduction in Kappa number. XECEDED-Refined bleached pulp resulted in handsheets with increased brightness compared to the control (X = xylanase treatment; E = alkaline extraction; C = Cl2 treatment; D = ClO2 treatment).     

Thermomyces lanuginosus: properties of strains and their hemicellulases

The non-cellulolytic Thermomyces lanuginosus is a widespread and frequently isolated thermophilic fungus. Several strains of this fungus have been reported to produce high levels of cellulase-free beta-xylanase both in shake-flask and bioreactor cultivations but intraspecies variability in terms of beta-xylanase production is apparent. Furthermore all strains produce low extracellular levels of other hemicellulases involved in hemicellulose hydrolysis. Crude and purified hemicellulases from this fungus are stable at high temperatures in the range of 50-80 degrees C and over a broad pH range (3-12). Various strains are reported to produce a single xylanase with molecular masses varying between 23 and 29 kDa and pI values between 3.7 and 4.1. The gene encoding the T. lanuginosus xylanase has been cloned and sequenced and is shown to be a member of family 11 glycosyl hydrolases. The crystal structure of the xylanase indicates that the enzyme consists of two beta-sheets and one alpha-helix and forms a rigid complex with the three central sugars of xyloheptaose whereas the peripheral sugars might assume different configurations thereby allowing branched xylan chains to be accepted. The presence of an extra disulfide bridge between the beta-strand and the alpha-helix, as well as to an increase in the density of charged residues throughout the xylanase might contribute to the thermostability. The ability of T. lanuginosus to produce high levels of cellulase-free thermostable xylanase has made the fungus an attractive source of thermostable xylanase with potential as a bleach-boosting agent in the pulp and paper industry and as an additive in the baking industry.     

Short Communication: Effect of agitation rate on the growth and production of xylanase free of cellulase by Thermomyces lanuginosus MH4 in bioreactor

Cellulase-free xylanase production by T. lanuginosus MH4 was investigated in a 3-litre stirred tank bioreactor under different agitation rates and an aeration rate of 1v/v/m. The cultivation time in the bioreactor was reduced significantly over that in shake culture conditions. A xylanase productivity of 0.1 mkat1–1h–1 was achieved on xylan in the bioreactor. This was nearly double to that obtained in shake culture. The agitation rates influenced both growth and enzyme secretion in the bioreactor. The highest level of biomass concentration and activities of both xylanase and -xylosidase were obtained at 150 revmin–1