一轮复习学案—课题：酵母细胞的固定化.doc

well, with the effectiveness of services to defend the interests of the masses. Third, we should strive to do well. To achieve good practical results, the key is to know the law, to grasp the laws and using laws. Office of economic development, social progress, there are rules to follow. Only act according to the law, to overcome blindness and strengthening initiative, creative. Working in the Office, we should be good at analyzing the essence of things, to find regular thing, change from passive to active, to seek one. Investee 2. establishment of evaluation system. Evaluation system is essentially an incentive mechanism, the evaluation was objective and fair, reasonable, and can stimulate a person's energy, mobilizing people's enthusiasm or be misleading, dampened the enthusiasm of people. At present, the concept of people-oriented people, but to establish and perfect evaluation system is still lagging behind. Work in this area should be seriously caught up. "Three emphasis on" to take advantage of. Is a focus on standards and scientific. A fundamental point of the evaluation criteria, is to keep contact, development, comprehensive eye evaluation of cadres. Office work, both "record" and "potential performance"; both subjective efforts, take another look at the objective condition; both "pragmatic" enough, take another look at the "retreat" level. Second, focus on participating the breadth of the subject. Evaluation of cadres must give full play to democracy, cadres and the masses handing over evaluation. Office work services leadership and service base, serve the people, then nominated the main object will contain a variety of services, so that assessment was accurate and to avoid one-sidedness. Third, pay attention to the seriousness of the conclusion. The use of evaluation findings is mainly embodied in his direction. Permitted to choose one, it would be tantamount to establishing a banner. Evaluation can't take care of the balance, otherwise it would undermine the offset the positive significance of evaluation. 　　Office of the party committees at all levels should take the findings as on the cadres ' bonus-penalty, an important basis for future movements, incentive and restraint effect of making evaluations. 3. Prevention of understanding "infidelity". People-centered development, concrete is dialectical,. In practice in the process, to prevent one-sided and still look, from actual people. One is to use dialectical point of view to understand. Humanist is the core of the scientific Outlook on development, remains the development of the scientific Outlook on development emphasizes, is still taking economic construction as the Center. Any departure from the development of thought and action, is a misinterpretation and misunderstanding of people-oriented. Adhere to people-oriented both to overcome one-sidedness of the old, but also to prevent one-sidedness, not accelerating the development of people and the opposition. Must be very clear, people oriented and speeding up development is not opposed, but consistent, we should never bestrengthened password centralized management, enhanced password communication network and the construction of electronic Government Affairs network, strengthen the translation of code cables work, strengthen the management of important files, make sure that the password is absolutely safe, ensure that Cryptography is absolutely clear. History of work to write a history of the original, compiled a book called changde Yearbook, out of a journal of the history of the publication, production of a song of the past after reading forums, changde, a story of people's hard work and exhibition. Archives to go the path of managing archives according to law, strengthen the construction of all types of archives, with the focus on key project archives, private enterprises, credit and Community Archives Archives, making full use of electronic means of technology and network, and promote the standardization and legalization of archives and information. Investee 4. insist on the logistics, will be around strong strong reforms, cadres of service management. Office workers is to be for this, deepen reform, strengthen management, and continuously meet the cadres of the organs function properly and the need for material and cultural life. One is to enhance the logistical support capability. Adapt to the new situation of the market economy, making full use of social power, flexibility in the use of market mechanisms and accelerate the process of marketization and socialization of logistic services. Adhere to the institutionalization of the logistics management, establish and implement a financial management system, trolley management systems, health management systems, security systems, such as the logistics management system, strengthen the management of human, financial, material, and logistical services standardized and institutionalized track. Secondly, we should strengthen the management of construction. Increasing investment, improving the working conditions of organs; overall civilization create an activity, and constantly improve the level of civilization; to promote greening, lighting, purification, flowers, landscaping, Office workers create a good environment for work and life. Third, to improve staff welfare. To in policy allows of range within, trying to for Office cadres workers solution housing, and couples separation, and children employment, aspects of actual difficult provides help, has conditions of place and units, also should appropriate to increased cadres workers of welfare, real do "career left people, feelings left people, appropriate treatment left people", let General Office staff in pay hard efforts of while, feel organization of warm, get due of affordable. Investee 5. insist on the team, will focus on stimulating vitality who care and respect for people. To Office cadres for the cut ... Learning style, strengthen the theoretical knowledge, optimize the structure of knowledge, and continuously improve the quality of cadres and workers. Attention to their physical health, and actively carry out regular wholesome recreational
【课 题】：课题4 酵母细胞的固定化
【复习目标】
1．说出固定化酶和固定化细胞的作用和原理。
2．尝试制备固定化酵母细胞，并利用固定化酵母细胞进行酒精发酵。
【复习重点与难点】
复习重点：制备固定化酵母细胞。
复习难点：制备固定化酵母细胞
【知识回顾】一、基础知识
酶：优点：催化效率高，低耗能、低污染，大规模地应用于食品、化工等各个领域。
实际问题：对环境条件敏感，易失活；溶液中的酶很难回收，不能再次利用，提高了生产成本；反应后的酶会混合在产物中，如不除去，会影响产品质量。
设想：
固定化酶：优点
实际问题：一种酶只能催化一种化学反应，而在生产实践中，很多产物的形成 都是通过一系列的酶促反应才能得到的
设想：
固定化细胞：优点
二、固定化酶的应用实例
高果糖浆是指
能将葡萄糖转化为果糖的酶是 。使用固定化酶技术，将这种酶固定在一种 上，
再将这些酶颗粒装到一个反应柱内，柱子底端装上分布着许多小孔的 。酶颗粒无法通过筛板的小孔，而反应溶液却可以自由出入。生产过程中，将葡萄糖溶液从反应柱的上端注入，使葡萄糖溶液流过反应柱，与 接触，转化成果糖，从反应柱的下端流出。反应柱能连续使用半年，大大降低了生产成本，提高了果糖的产量和质量。
三、固定化细胞技术
固定化酶和固定化细胞是利用 或 方法将酶或细胞固定在一定空间内的技术，包括 、 和 法。
一般来说，酶更适合采用 和 法固定，而细胞多采用 法固定化。这是因为细胞个大，而酶分子很小；个大的细胞难以被 或 ，而个小的酶容易从 中漏出。
包埋法法固定化细胞即将微生物细胞 包埋在不溶于水的 中。常用的载体有 、 、 、 和 等。
四、实验操作
(一)制备固定化酵母细胞
制备固定化酵母细胞需要的材料是 、 、 、 、
、 、 、 、 和
1.酵母菌的活化
活化就是处于 状态的微生物重新恢复正常的生活状态。
2.配制物质的量尝试为0.05mol/L的CaCl2溶液
3.配制海藻酸钠溶液
加热溶化海藻酸钠时要注意：

4.海藻酸钠溶液与酵母菌细胞混合
5.固定化酵母细胞
以恒定的速度缓慢地将注射器中的溶液滴加到配制好的 溶液中，并浸泡 (时间)。
(二)用固定化酵母细胞发酵
1.将固定好的酵母细胞用 冲洗2-3次。
2.发酵时的温度就为 ，时间 。
五、结果分析与评价
（一）观察凝胶珠的颜色和形状
如果制作的凝胶珠颜色过浅、呈白色，说明 ；如果形成的凝胶珠不是圆形或椭圆形，则说明 ，制作失败，需要再作尝试。
（二）观察发酵的葡萄糖溶液
利用固定的酵母细胞发酵产生酒精，可以看到产生了很多 ，同时会闻到
补充：直接使用酶、固定化酶和固定化细胞催化的优缺点：
类型
优点
不足
直接使用酶
催化效率高，低耗能、低污染等。
对环境条件非常敏感，容易失活；溶液中的酶很难回收，不能被再次利用，提高了生产成本；反应后酶会混在产物中，可能影响产品质量。
固定化酶
酶既能与反应物接触，又能与产物分离，同时，固定在载体上的酶还可以被反复利用。
一种酶只能催化一种化学反应，而在生产实践中，很多产物的形成都通过一系列的酶促反应才能得到的。
固定化细胞
成本低，操作更容易。
固定后的酶或细胞与反应物不容易接近，可能导致反应效果下降等。

注意事项
1.配制海藻酸钠溶液：小火、间断加热、定容，如果加热太快，海藻酸钠会发生焦糊。
2.海藻酸钠溶液与酶母细胞混合：冷却后再混合，注意混合均匀，不要进入气泡
3.制备固定化酵母细胞：高度适宜，并匀速滴入
4．刚形成的凝胶珠应在CaCL2溶液中浸泡一段时间，以便Ca2+与Na+充分交换，形成的凝胶珠稳定。检验凝胶珠是否形成，可用下列方法：用镊子夹起一个凝胶珠放在实验桌上用手挤压，如果不容易破裂，没有液体流出就表明成功地制成了凝胶珠，还可以用手将凝胶珠在实验桌上用力摔打，如果凝胶珠很容易弹起，也表明制备的凝胶珠是成功的。
5．凝胶珠的颜色和形状
如果制作的凝胶珠颜色过浅、呈白色，说明海藻酸钠的浓度偏低，固定的酵母细胞数目较少；如果形成的凝胶珠不是圆形或椭圆形，则说明海藻酸钠的浓度偏高，制作失败，需要再作尝试。
【典例解析】
1．酶和细胞分别适应哪种固定方法？

【课堂练习】
1．下列关于固定化酶中用到的反应柱的理解中。正确的是 ( )
A．反应物和酶均可自由通过反应柱 B．反应物和酶均不能通过反应柱
C．反应物能通过，酶不能通过 D．反应物不能通过，酶能够通过
2.如果反应物是大分子物质，采用那种方法催化受限制( )。
A．直接使用酶 B．使用化学方法结合的酶
C．使用固定化细胞 D．使用物理吸附法固定的酶
3．固定化细胞常用包埋法固定，原因是( )
A．包埋法固定操作最简便 B．包埋法对酶活性的影响最小
C．包埋法固定具有普遍性 D．细胞体积大，难以被吸附或结合
4．下列不属于固定化酶在利用时的特点的是 ( )
A．一种固定化酶一般不能催化一系列酶促反应B．有利于酶与产物分离，提高产物品质
C．能自由出入依附的载体D．酶是催化剂，但固定化酶也不可永远利用
5.固定化细胞对酶的活性影响最小，根本原因是( )。
A．避免了细胞破碎、酶的提取纯化过程 B．固定化细胞在多种酶促反应中连续发挥作用
C．促化反应结束后，能被吸收和重复利用
D．细胞结构保证了各种酶在细胞内化学反应中有效的发挥作用
6．下列关于固定化酶和一般酶制剂在应用效果上的说法中，错误的是 ( )
A．固定化酶生物活性强，可长久使用
B．一般酶制剂应用后和产物混在一起，产物的纯度不高
C．一般酶制剂只能一次性利用 D．固定化酶可降低生产成本，提高产量和和质量
7、 使用固定化酶的优点是 ( )
A．有利于增加酶的活性 B．有利于产物的纯化
C．有利于提高反应速度 D．有利于酶发挥作用
8.下列关于酶和细胞的固定叙述不正确的是( )。
A． 酶分子很小，易采用包埋法
B．酶分子很小，易采用化学结合或物理吸附法固定的酶
C．细胞个大，难被吸附或结合 D．细胞易采用包埋法固定
9．下列不是用于包埋法固定化细胞的载体是( )
A．琼脂糖 B．醋酸纤维素 C．聚丙烯酰胺 D聚乙烯树脂
10．下列关于固定化细胞的叙述，正确的是 ( )
A．催化效率高，低耗能、低污染 B．对环境的条件非常敏感，容易失活 C．既能与反应物充分接触，又能与产物分离 D．成本低、操作简单，但反应效率可能很低
11．溶解海藻酸钠，最好采用下列哪种加热的方法 ( )
A．小火间断 B．小火持续 C．大火间断 D．大火持续
12．在制备固定化酵母细胞的实验中，CaCl2溶液的作用是 ( )
A．调节溶液pH B．进行离子交换
C．胶体聚沉 D．为酵母菌提供钙离子

13.下图为固定化酶的反应柱示意图，请据图回答：
(1)请请在横线上填出各标号名称.①________
②_________ ③_________。
(2) 与一般酶制剂相比，固定化酶的突出优点
是_________________________________
和______________________
固定化细胞的优点是___________________________________。
(3)③的作用是___________________________________________________________。
（4）制作固定葡萄糖异构酶所用的方式有___________________或_________________________。
（5）据图说出反应柱的作用原理
。
14.下图是制备固定化酵母细胞实验步骤图解，请据图回答：
酵母细胞活化→配制CaCl2溶液→配置海藻酸钠溶液→海藻酸钠溶液与酵母细胞混合→固定化酵母细胞
(1) 在_______________状态下，微生物处于休眠状态。活化就是让处于休眠状态的微生物恢复_________状态。活化前应选择足够大的容器，因为酵母细胞活化时_________。
(2) 影响实验成败的关键步骤是_______________ 。
(3) 如果海藻酸钠浓度过低，形成凝胶珠包埋酵母细胞数目______。
（4）观察形成凝胶珠的颜色和形状，如果颜色过浅，说明
如果形成的凝胶珠不是圆形或椭圆形，说明 。
（5）制备固定化酶不宜用包埋法，为什么？

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15、答案：（1）反应柱； 固定化酶； 分布着小孔的筛板 （2）既能与反应物接触，又能与反应物分离； 能被反复利用； 成本更低，操作更容易 （3）酶颗粒无法通过筛板上的小孔，反应溶液可自由出入。（4）将酶相互连接起来；将酶吸附在载体表面上 （5）反应物溶液从反应柱上端注入，使反应物溶液流过反应柱，与固定化酶接触，得到产物，从反应柱下端流出。

16、答案：（1）缺水； 正常生活状态； 体积增大（2）配制海藻酸钠溶液； (3)少
（4）固定化酵母细胞数目较少； 海藻酸钠浓度偏高，制作失败
（5）因为酶分子很小，容易从包埋材料中露出

对需取得患者书面同意方可进行的医疗活动，应当由患者本人签署知情同意书。患者不具备完全民事行为能力时，应当由其法定代理人签字；患者因病无法签字时，应当由其授权的人员签字people-oriented in the name of reducing our enthusiasm. Premier Wen Jiabao's Government work report this year, identified as 7% growth, 2.1% lower than last year's 9.1%, but in 8 work remains to accelerate development in the first place ... Work standards. Second is to use the scale of development to understand. Society is evolving, human needs are also changing. Practice of humanism is a long-term process and cannot be achieved overnight. To treat people if you stand still and rigid dogma, it is possible to return to this, see things but not people's old ways. Therefore, we have to follow the development of the times, and continue debugging the humanist perspective, innovative people-oriented way. Under the new historical conditions, we are requested by the times, set up a brand new concept, a high starting point, high standard of armed people, shaping, exercise, people, constantly taking the new connotation of the times. Terms of Office, is to lead the vast numbers of cadres to actively adapt to the knowledge faster and actively adapt to the increasingly high level of leadership situation, actively adapt to the demand of the masses more and more situations, eager to learn, enhance accumulation, efforts, honed the ability to overcome a good service, panic. Third is to use realistic understanding. Adhere to people-oriented must be based on reality, not divorced from reality. Desk no water cups here in Hong Kong, staff need to drink some water to the drinking fountain on a floor, the glass used is a one-time non-cone type. Drink this cup can only hold, cannot be put down. This is not only to save on glass materials, more important is to force you to get drinking water, go back to your seat business. 　　Our emphasis on people-oriented, does not imply you can indulge the desires, the comfortable, the pursuit of enjoyment. Office of the party committees at all levels of our comrades, must continue to maintain the style of plain living and hard struggle, to stand up to the tests of temptations, exciting "energy" conservation "disposition" to the spirit of selfless dedication and provide services. Investee 4. clearly defined responsibilities. Putting people first is a common vision of development, everyone is a liability. People-oriented, above and below with the responsibility for coordinating system must be established and hold everyone accountable. First, we must strengthen the leadership. "In its place, its governance, its level, bear the responsibility of the Division." This is the minimum requirement for leaders, is a measure of the basic criteria for whether a leading cadre is competent. Leaders need to "it is an official term for the benefit of" mission, consciously assume responsibility for implementing people-oriented. Director of the Office of the party committees at all levels is the responsibility of the units to implement people-oriented, to manage their people, "plug in own responsibility", effectively enhance the education of leading cadres, cadres and cadres work, efforts to revitalize the Office of human resources, Office work is full of vigor and vitalityactivity, and constantly improve the physical quality of cadres and workers. Broaden the channels into his Office, increase the intensity of writing the open selection of cadres and cadres there, change text structure is not optimal conditions, and continuously improve the overall quality of cadres and workers. The second is to create a relaxed environment. With a deep affection for Office cadres, cadres and the mentality of the concern, attention to their conditions of l