己酸乙酯酯化真菌筛选及发酵条件研究

本文主要研究了具有己酸乙酯酯化活性的真菌菌株的筛选和发酵条件优化。从大曲和糟醅样品中分离纯化获得79株产生透明圈的丝状真菌，菌落形态初步识别结果显示分离菌株包括红曲霉属、根霉属及曲霉属等菌株。其中菌株EM-56酯化酶活力最强，发酵获得的粗酶制剂酶活为172.36 u。根据显微形态、菌落形态及生理生化特征，初步鉴定该菌株为曲霉科红曲霉属紫色红曲霉（Monascus purpureus）。 在此基础上重点研究了菌株EM-56在不同培养基成分及不同培养条件下的产酶情况，确定了最佳培养基和培养条件。通过单因素实验确定在基础培养基中添加最佳碳源为葡萄糖，最佳氮源为蛋白胨。正交优化实验结果确定了最佳培养基组成：以麸皮为基础培养基，添加葡萄糖 2%，蛋白胨 0.3 %，KH2PO4.3H2O 0.05 %，MgSO4.7H2O 0.06 %。菌株EM-56在上述培养基中的最佳发酵条件为：初始pH 5.5，发酵温度为35°C，发酵时间7d，种龄48h，接种量8%，装瓶量50g / 瓶（500mL）。在最佳培养基和发酵条件下，菌株EM-56发酵获得的粗酶制剂酶活达到241.56 u，比优化前提高了40.15%。 In this paper, the research focuses on the selection of fungus with esterifying activity and optimization of fermentation conditions. We isolated 79 strains which had transparent zones from Daqu and fermented grains. The isolated strains contained Monascus、Rhizopus and Aspergillus through primary morphology analysis. The strain of EM-56 which produces strongest esterase was selected. The enzyme activity reached 172.36u. According to related literature, EM-56 was identified as Monascus purpureus through morphology analysis and biochemical determination. We also studied the effects of different medium and fermentation conditions on the esterase production of strain EM-56. The optimal medium and fermentation conditions were determined. Single factor experiment result shows that the optimal carbon source added is glucose and the optimal nitrogen source added is peptone. The optimal fermentation medium determined by orthogonal optimization test is as follows: wheat bran as substrate, glucose 2%, peptone 0.3%, KH2PO4.3H2O 0.05%，MgSO4.7H2O 0.06%. The optimal fermentation conditions are: initial pH 5.5, cultural temperature 35°C, cultural time 7d, seed age 48h, inoculation 8%, medium mass 50g / flask（500mL）. The esterse activity of EM-56 cultivated in the optimal medium and fermentation conditions reached 241.56u and increased by 40.15% compared with the original activity.

两株耐碱性木聚糖酶产生菌的筛选及产酶条件研究

本文主要研究了从造纸厂碱性土壤中筛选得到的，能够产生耐碱木聚糖酶的两株放线菌X24-14和X15-17。通过16 S rRNA基因序列分析并结合菌株的形态特征以及生理生化特性，初步认为菌株X15-17为拟诺卡氏菌属（Nocardiopsis）的一个潜在新种；菌株X24-14为纤维化纤维菌（Cellulosimicrobium cellulans）。 在此基础上探索了菌株X24-14和菌株X15-17所产木聚糖酶的基本酶学性质。研究发现，两株菌所产的木聚糖酶的耐碱性均较强： 1）菌株X24-14所产的木聚糖酶，在pH 4.2～9.4的范围内能维持较高的活力，pH 9.4条件下，仍能保持80％的酶活力；2）菌株X15-17所产的木聚糖酶在pH 4.0～9.0的范围内能维持较高的活力，pH 9.0条件下，仍能保持80％的酶活力；3）两株菌所产的木聚糖酶均具有较好的pH稳定性，在pH 2.0～11.0范围内稳定，pH 11.0、4 ℃条件下处理24 h仍具有75％的活力。 本文还重点研究了菌株X24-14在不同培养基成分及不同培养条件下的产酶情况，确定了其适宜的产酶条件。结果显示，菌株X24-14的最适碳源为麸皮；最适氮源为蛋白胨；最适产酶pH为pH 8.5。菌株X24-14适宜的产酶条件为：麸皮60 g/L，蛋白胨10 g/L，K2HPO4 7.0 g/L，pH 8.5，接种量为5％，37 ℃，200 r/min发酵培养108 h。 Two strains of actinomycetes, X24-14 and X15-17, which produced alkali-tolerant xylanase were screened from the soil samples collected from a pulp mill in china. Based on the morphological, physiochemical characteristics and 16S rRNA sequence, X24-14 was priminarily identified as cellulosimicrobium cellulans ; X15-17 was priminarily identified as a new species of Nocardiopsis. The investigation examined the enzyme activities which produced by X24-14 and X15-17 under different pH and different temperatures. The results showed that : 1）The xylanase from X24-14 had characteristic of alkali-tolerance: It remains 80% relative activity at pH ranges between pH 4.2 and pH 9.4 under 50℃. 2)The xylanase from X15-17 also showed characteristic of alkali-tolerance, it remains 80% relative activity at pH ranges between pH 4.0and pH 9.0 under 50℃. 3）The xylanase from the two strains showed alkali-stable characteristics. They were stable at pH ranges between pH 2.0 and pH 11.0, showing 75% of its maximal activity remaining under 24 hours of treatment at 4℃. We also studied the effect of different growth conditions: carbon source, nitrogen sources, inoculum size, and initial pH on the production of xylanase of strain X24-14. The results showed that :The optimal carbon source was wheat bran; The optima nitrogen source was peptone; The maximum xylanase activity was achieved in the medium containing 60 g/L wheat bran, 10 g/L peptone, 7 g/L K2HPO4, inoculum size 5% and pH 8.5, under 37℃ in 108 h.

准噶尔盆地中拐东北斜坡油气成藏条件研究——西部压性盆地含油气系统理论的研究与实践

This paper builds the model of oil accumulation and achieves the prediction of exploration goal. It uses multiple subject means, the ways of synthetic research and the viewpoint of analyzing genesis, with the academic guidance of sedimentology, structural geology, petroleum geology and geochemistry, the basis of strata sequence frame and structural frame, the frame of "four history" - the burying history, the structural history, the filling history and the evolving history of oil, the masterstroke of hydrocarbon's generation, migration and accumulation, the aim of revealing the genetic relation between mature source rock and oil reservoir in space and time. Some achievements and viewpoints in this study are following. 1. It is proposed that the structural evolution in this area had many periods, and the structural movement of the Xiazijie group telophase formed the structural pattern for the first time. 2. The character of strata sequence in this area is divided by the character of episodic cycle firstly. The study of dividing the facies of single well and the facies of well tie is based on the data of single well. The character of sedimentary facies is con-structed initially. 3. It is believed that Jiamuhe group is the main source rock, which can supply considerable oil and gas resources for the first time. Some criterions of source rock such as the type ,the abundance in Jiamuhe group are analysed. Using the thermal history of source rock, we drawn a conclusion that the original type of source rock in Jiamuhe group is II_1-III, and the abundance achived the level of good source rock, and this set of source rock had contributed to this area. 4. The reservoir strata in this area are assessed and analysed with the reservoir evaluation. There are multi-type reservoirs, such as volcanic lava facies, sedimentary clast facies, continental belch facies. The physical property in reservoir strata is characterized by low porosity and low permeability. The study of diagenetic stage show that the diageneses in Jiamuhe group is A-Bsubage, and the reservoir room is mainly secondary corroded hollow and cleft. 5. The synthetic research on oil system in Jiamuhe group is made for the first time. The type of petroleum system is divided , and we consider that the petroleum system of Jiamuhe group is at the reliable rank. There are two critical time in oil accumulation through studying the critical time of oil accumulation : the early generation of hydrocarbon is oil, and the later is gas. 6. The mechanism of accumulation is analysed. We consider that the accu-mulation of oil in this area has many periods, and the early generated hydrocarbon is expeled by the later , and formed the character of zonal distribution in planar. 7. A bran-new model of oil and gas is proposed. Beneficial enrichment area of oil and gas is analyzed, which can be divided into three sections: Section I can be divided into two sections: I_1 and I_2. The lower subgroup of Jiamuhe is covered by the triassic layer of I_1 section. Fault zone and near the foot wall of fault are charactered with thick phase belt. Then the cover capability in this area is relatively poor, oil can migrate into triassic layer by vertical or lateral migration , and forms I_1 Kelamayi triassic oil pool consequently. The lower subgroup of Jiamuhe is covered by the triassic layer of I_2 section ,which is charactered with thin phase belt. Then the cover capability in this area is relatively good, and forms I_1 Kelamayi triassic oil pool consequently. Section II can be divided into two sections: II_1-I_(I~2). The cover of Jiamuhe group in section II_1 is the low resistivity segment in Wuerhe group, which has thin lithology and poor porosity and permeability. Oil and gas in Jiamuhe group can be covered to form beneficial accumulation area. There are some wells in this area, such as Ke 007 well, 561 well. The thick phase belt layer of Wuerhe high resistivity segment in section II_2 has unconformable relation with Jiamuhe group. The cover ability of the high resistivity segment is poor, petroleum in Jiamuhe can migrate into Wuerhe layer vertically. This area is the beneficial area for accumulating petroleum in Wuerhe layer. there are some wells in this area, such as Ke 75 well, Ke 76 well, Ke 77 well, Ke 78 well, Ke 79 well. Section III can also be divided into two sections: III_1 and III_2. Wuerhe group in section III_1 has unconformable relation with Jiamuhe group. There is thick lithology and poor cover in Wuerhe group, but the strata sequence evolution character of upper subgroup in Jiamuhe group has determined that it has lateral and vertical cover ability. thus, this area is petroleum abundant belt of jiamuhe group, which has the trap. Section III_2 is an area controled by wedgeout of Fengcheng group, Fengcheng group in this area has quite thick lithology so that It has beneficial resevoir phase belt. It can accumulate oil in itself or accept some oil in Jiamuhe group. Jiamuhe group has some oil accumulation condition in this area. Thus, section III_2 is jiamuhe-Fengcheng multiple petroleum accumulation belt, such as Ke 80 well. 8. The goal of exploration is suggested: Depositional trap or combination trap is the important aspect in later exploration. Both types of traps are the goal of the next drilling: Fault block trap in the east of 576 well and the NO. 2 fault block trap in the north of Ke 102 well It is suggested that we should study the law of oil and gas in Jiamuhe group and enhance the study of combination in forming reservoir and trap scale. We do some lithology forecast and reservoir diatropic forecast in order to know the area of oil and gas.