📚 Enzymes | 酶
Enzymes are biological catalysts that accelerate the rate of metabolic reactions without being consumed in the process. They are predominantly globular proteins with highly specific active sites, allowing them to control virtually every biochemical reaction in living organisms. Understanding enzyme structure, kinetics, and regulation is fundamental to A‑Level Biology, and the CCEA specification places particular emphasis on the induced‑fit model, factors affecting enzyme activity, and the distinction between competitive and non‑competitive inhibition.
酶是生物催化剂,能在不被消耗的情况下加速代谢反应的速率。它们大多为球状蛋白质,拥有高度特异性的活性部位,能够控制生物体内几乎所有的生化反应。理解酶的结构、动力学和调控是A‑Level生物的基础,CCEA 考试大纲特别强调诱导契合模型、影响酶活性的因素以及竞争性抑制剂与非竞争性抑制剂的区别。
1. What are Enzymes? | 什么是酶?
Enzymes are globular proteins that function as biological catalysts. They lower the activation energy of a reaction, enabling metabolic processes to occur rapidly at body temperature. Without enzymes, most cellular reactions would be too slow to sustain life. Each enzyme is specific to a particular substrate or a group of related substrates, and their names often end in ‘‑ase’, such as amylase, protease, and catalase.
酶是球状蛋白质,作为生物催化剂发挥作用。它们降低反应的活化能,使代谢过程能够在体温下快速进行。如果没有酶,大多数细胞反应将过于缓慢而无法维持生命。每种酶对特定的底物或一组相关底物具有特异性,其名称通常以“‑ase”结尾,如淀粉酶(amylase)、蛋白酶(protease)和过氧化氢酶(catalase)。
Enzymes are not altered by the reaction they catalyse, meaning a single enzyme molecule can be reused many times. Some enzymes are RNA‑based (ribozymes), but the syllabus focuses exclusively on protein enzymes. The catalytic power of enzymes is immense; they can increase reaction rates by factors of 10⁶ to 10¹² compared with the uncatalysed reaction.
酶不会因其催化的反应而改变,这意味着一个酶分子可以被重复使用多次。有些酶是RNA构成的(核酶),但课程大纲只关注蛋白质酶。酶的催化能力极其强大,与无催化的反应相比,它们能将反应速率提高10⁶到10¹²倍。
2. Structure of Enzymes | 酶的结构
Enzymes are large proteins folded into a precise three‑dimensional shape. The sequence of amino acids (primary structure) dictates how the polypeptide chain coils (secondary structure) and folds into a compact globular conformation (tertiary structure). The active site is a cleft or pocket formed by a specific arrangement of amino acid residues. This unique shape allows the enzyme to bind to its substrate with high specificity.
酶是折叠成精确三维形状的大分子蛋白质。氨基酸序列(一级结构)决定了多肽链如何盘绕(二级结构)并折叠成紧密的球状构象(三级结构)。活性部位是由特定氨基酸残基排列形成的裂隙或口袋,这种独特的形状使酶能够以高度特异性结合其底物。
Some enzymes consist only of amino acids, while others require a non‑protein component called a cofactor to be active. Cofactors can be inorganic ions (e.g. Zn²⁺, Fe²⁺) or complex organic molecules called coenzymes (e.g. NAD⁺, FAD, coenzyme A). The protein part of such an enzyme is called the apoenzyme, and the complete, catalytically active complex is the holoenzyme.
有些酶仅由氨基酸组成,而另一些则需要一种称为辅因子的非蛋白质成分才能具有活性。辅因子可以是无机离子(如 Zn²⁺、Fe²⁺),也可以是称为辅酶的复杂有机分子(如 NAD⁺、FAD、辅酶A)。这类酶的蛋白质部分称为脱辅酶,完整的具有催化活性的复合物称为全酶。
3. Mechanism of Enzyme Action | 酶的作用机制
The function of an enzyme is explained by the formation of an enzyme‑substrate complex. The classic lock‑and‑key model suggests that the active site has a fixed, rigid shape perfectly complementary to the substrate, much like a key fitting a lock. While this model illustrates specificity, it cannot explain how enzymes stabilise the transition state or why some molecules with similar shape can inhibit activity.
酶的功能可通过酶‑底物复合物的形成来解释。经典的锁钥模型认为,活性部位具有固定的、刚性的形状,与底物完全互补,就像钥匙插入锁孔一样。虽然该模型能说明特异性,但无法解释酶如何稳定过渡态,也无法解释为何形状相似的一些分子能够抑制酶的活性。
The induced‑fit model, accepted today, proposes that the active site is flexible. When a substrate enters, the binding induces a conformational change in the enzyme, causing the active site to wrap around the substrate and mould itself into a precise fit. This change puts strain on chemical bonds in the substrate, lowering the activation energy and facilitating the transition state. This model better accounts for the catalytic efficiency and regulation of enzymes.
今天被广泛接受的诱导契合模型认为,活性部位是柔性的。当底物进入时,结合会诱导酶发生构象改变,使活性部位包绕底物,并自身微调以达到精确契合。这种变化使底物中的化学键承受张力,从而降低活化能并促进过渡态的形成。该模型能更好地解释酶的催化效率和调节作用。
4. Activation Energy | 活化能
All chemical reactions involve an energy barrier known as the activation energy (Eₐ) — the minimum energy required for reactants to collide successfully and form products. Enzymes work by lowering this activation energy. They do so by providing an alternative reaction pathway with a lower energy transition state, often through the strain, proximity, and orientation effects created within the enzyme‑substrate complex.
所有化学反应都涉及一个称为活化能(Eₐ)的能量壁垒——即反应物成功碰撞并形成产物所需的最低能量。酶通过降低活化能来发挥作用。它们通过提供一条具有较低能量过渡态的替代反应路径来实现,通常借助酶‑底物复合物中产生的张力、靠近和定向效应。
On an energy profile diagram, the uncatalysed reaction has a high peak, whereas the enzyme‑catalysed reaction shows a much lower peak. The overall free‑energy change (ΔG) of the reaction remains the same; enzymes do not alter the equilibrium position or the nature of the products, only the speed at which equilibrium is reached.
在能量变化曲线图中,无催化反应具有较高的峰,而酶催化反应的峰则低得多。反应的总自由能变化(ΔG)保持不变;酶不改变平衡位置或产物的性质,只改变达到平衡的速度。
5. Factors Affecting Enzyme Activity: Temperature | 影响酶活性的因素:温度
Enzyme activity increases with temperature up to an optimum point, typically around 37 °C for human enzymes. The rise in kinetic energy speeds up molecular movement, increasing the frequency of successful collisions between enzyme and substrate. However, beyond the optimum temperature, the rate declines sharply as the enzyme begins to denature. Denaturation involves the disruption of hydrogen bonds, ionic bonds, and hydrophobic interactions that maintain the tertiary structure, causing the active site to lose its shape permanently.
酶活性随着温度升高而增加,直至达到最适温度,人类酶的最适温度通常在37°C左右。动能的增加加快了分子运动,提高了酶与底物成功碰撞的频率。然而,超过最适温度后,反应速率急剧下降,因为酶开始变性。变性会破坏维持三级结构的氢键、离子键和疏水相互作用,导致活性部位永久变形。
The temperature coefficient Q₁₀ describes how the rate roughly doubles for every 10 °C rise within the physiological range. In thermophilic bacteria, enzymes have evolved to withstand temperatures of 70 °C or higher without denaturation, making them valuable in industrial biotechnology. Students should be able to sketch and interpret the typical bell‑shaped temperature–activity curve.
温度系数 Q₁₀ 描述了在生理范围内,温度每升高10°C,反应速率大致加倍的现象。在嗜热细菌中,酶已进化到能耐受70°C甚至更高的温度而不变性,这使得它们在工业生物技术中极具价值。学生应能够绘制并解释典型的钟形温度‑活性曲线。
6. Factors Affecting Enzyme Activity: pH | 影响酶活性的因素:pH
Each enzyme has a narrow optimum pH range. For example, pepsin in the stomach functions best at pH 2, while trypsin in the small intestine works optimally at pH 8. Changes in pH alter the ionisation state of amino acid side chains at the active site and throughout the enzyme. This can affect substrate binding, catalytic groups, and the overall conformation. Like extreme temperature, extreme pH can cause irreversible denaturation.
每种酶都有狭窄的最适pH范围。例如,胃蛋白酶(pepsin)在pH 2时功能最佳,而小肠中的胰蛋白酶(trypsin)在pH 8时效率最高。pH的变化会改变活性部位及整个酶中氨基酸侧链的电离状态,从而影响底物结合、催化基团和整体构象。与极端温度类似,极端pH也会导致不可逆的变性。
The pH–activity curve is typically a symmetrical bell shape around the optimum. Buffer solutions are often used in laboratory experiments to maintain a constant pH when investigating enzyme activity, ensuring any observed changes are due solely to the intended variable. Students should understand the importance of buffers in both experimental design and in biological systems such as the blood.
pH‑活性曲线通常是以最适pH为中心的对称钟形曲线。在进行酶活性实验时,常使用缓冲溶液来维持恒定的pH,以确保观察到的任何变化完全由目标变量引起。学生应理解缓冲液在实验设计和血液等生物系统中的重要性。
7. Factors Affecting Enzyme Activity: Substrate Concentration | 影响酶活性的因素:底物浓度
At low substrate concentrations, the rate of reaction increases almost linearly with substrate concentration because many active sites are vacant. As substrate concentration continues to rise, the increase in rate becomes progressively smaller because an increasing proportion of active sites become occupied. Eventually, at high substrate concentration, the rate reaches a maximum velocity (Vₘₐₓ), where all active sites are saturated and the enzyme is working at its full capacity.
在底物浓度较低时,反应速率几乎随底物浓度线性增加,因为许多活性部位处于空闲状态。随着底物浓度继续上升,速率的增幅逐渐减小,因为越来越多的活性部位被占据。最终,在高底物浓度下,速率达到最大值(Vₘₐₓ),此时所有活性部位均已饱和,酶全力工作。
The relationship between substrate concentration and rate follows Michaelis–Menten kinetics, described by the equation:
V = (Vₘₐₓ × [S]) / (Kₘ + [S])
Kₘ, the Michaelis constant, is the substrate concentration at which the reaction rate is half of Vₘₐₓ. A low Kₘ indicates high affinity of the enzyme for its substrate, and vice versa. While full derivations are not required, CCEA candidates should be able to interpret Vₘₐₓ and Kₘ values from substrate concentration–rate graphs and understand how inhibitors affect these parameters.
底物浓度与速率之间的关系遵循米氏动力学,由以下方程描述:
V = (Vₘₐₓ × [S]) / (Kₘ + [S])
米氏常数 Kₘ 是反应速率达到 Vₘₐₓ 一半时的底物浓度。Kₘ 值低表明酶对底物的亲和力高,反之亦然。尽管不需要详细推导,CCEA 考生应能够从底物浓度‑速率图中解读 Vₘₐₓ 和 Kₘ 值,并理解抑制剂如何影响这些参数。
8. Enzyme Inhibition: Competitive and Non‑competitive | 酶抑制剂:竞争性与非竞争性
Inhibitors are substances that reduce enzyme activity. Competitive inhibitors have a structure similar to the substrate and compete for the active site. They can be overcome by increasing the substrate concentration. In the presence of a competitive inhibitor, Vₘₐₓ remains the same, but the apparent Kₘ increases, meaning a higher substrate concentration is needed to reach half‑maximum velocity.
抑制剂是降低酶活性的物质。竞争性抑制剂的结构与底物相似,与底物竞争活性部位。通过增加底物浓度可以克服其抑制作用。在竞争性抑制剂存在的情况下,Vₘₐₓ 保持不变,但表观 Kₘ 增大,意味着需要更高的底物浓度才能达到半最大速率。
Non‑competitive inhibitors bind to a site other than the active site (an allosteric site), altering the enzyme’s shape so that the active site is no longer functional. This type of inhibition cannot be overcome by adding more substrate. Vₘₐₓ decreases because the total number of functional enzyme molecules is reduced, while Kₘ remains unchanged because the uninhibited enzyme molecules still bind substrate with the same affinity.
非竞争性抑制剂结合于活性部位以外的位点(变构位点),改变酶的形状,使活性部位不再正常运作。这种抑制作用无法通过增加底物来克服。Vₘₐₓ 降低,因为有功能的酶分子总数减少,而 Kₘ 保持不变,因为未被抑制的酶分子仍以相同的亲和力结合底物。
Another important category is uncompetitive inhibition, where the inhibitor binds only to the enzyme‑substrate complex, but this is less emphasised. Examples of inhibitors include statins (competitive inhibitor of HMG‑CoA reductase) and cyanide (non‑competitive inhibitor of cytochrome c oxidase). The specificity of inhibitors makes them powerful tools in medicine and biochemistry.
另一个重要的类别是反竞争性抑制,抑制剂仅与酶‑底物复合物结合,但这一点强调较少。抑制剂的例子包括他汀类药物(HMG‑CoA 还原酶的竞争性抑制剂)和氰化物(细胞色素 c 氧化酶的非竞争性抑制剂)。抑制剂的专一性使其成为医学和生物化学中的强大工具。
9. Cofactors and Coenzymes | 辅因子与辅酶
Many enzymes require additional non‑protein chemical components for catalytic activity. Inorganic cofactors include metal ions such as Mg²⁺ for DNA polymerase and Fe²⁺ in catalase. Organic cofactors, or coenzymes, are often derived from vitamins; for instance, NAD⁺ is derived from niacin (vitamin B₃), and FAD is derived from riboflavin (vitamin B₂).
许多酶需要额外的非蛋白质化学成分才能发挥催化活性。无机辅因子包括金属离子,如 DNA 聚合酶所需的 Mg²⁺,过氧化氢酶中的 Fe²⁺。有机辅因子即辅酶,通常来源于维生素;例如 NAD⁺ 来源于烟酸(维生素 B₃),FAD 来源于核黄素(维生素 B₂)。
Coenzymes act as carriers of electrons, atoms, or functional groups between reactions. They are not permanently bound and may be released and reused in subsequent catalytic cycles. A prosthetic group is a cofactor that is tightly or covalently attached to the enzyme, such as the haem group in haemoglobin (though haemoglobin is not an enzyme) or the FAD in succinate dehydrogenase. The requirement for coenzymes highlights the link between diet and metabolism.
辅酶在反应之间充当电子、原子或官能团的载体。它们并非永久结合,可以被释放并在后续催化循环中被重复利用。辅基是紧密结合或共价连接在酶上的辅因子,例如血红蛋白中的血红素基团(尽管血红蛋白不是酶)或琥珀酸脱氢酶中的 FAD。对辅酶的需求突显了饮食与代谢之间的联系。
10. Immobilised Enzymes | 固定化酶
In industrial processes, enzymes are often immobilised — attached to an inert, insoluble material such as alginate beads, silica gel, or cellulose. Immobilisation provides several advantages: the enzyme can be easily recovered and reused, product contamination is minimised, and the enzyme’s stability is often improved, allowing it to tolerate a wider range of pH and temperature.
在工业过程中,酶常被固定化——附着在惰性、不溶的材料上,如藻酸盐珠、硅胶或纤维素。固定化具有多项优势:酶可以轻松回收并重复利用,最大程度减少产物污染,而且酶的稳定性往往得到提高,使其能耐受更宽的 pH 和温度范围。
Common methods of immobilisation include physical entrapment within a gel matrix, adsorption onto a solid surface, covalent bonding to a support, and membrane confinement. An example is the use of immobilised lactose‑free milk, where lactase bound to beads hydrolyses lactose into glucose and galactose, providing a commercially viable solution for lactose‑intolerant individuals.
常见的固定化方法包括在凝胶基质中的物理包埋、吸附在固体表面、与载体共价结合以及膜限域。一个例子是使用固定化乳糖酶生产无乳糖牛奶,其中结合在珠粒上的乳糖酶将乳糖水解为葡萄糖和半乳糖,为乳糖不耐受人群提供了一种商业上可行的解决方案。
11. Industrial and Medical Applications of Enzymes | 酶的工业与医学应用
Enzymes are used across numerous industries. In food processing, pectinases clarify fruit juices, and proteases tenderise meat. In detergent manufacturing, lipases and cellulases improve stain removal. In medical diagnostics, enzymes like glucose oxidase are used in biosensors to measure blood glucose concentrations. Enzyme‑linked immunosorbent assays (ELISAs) rely on enzyme‑antibody conjugates to detect specific antigens or antibodies, such as in HIV testing.
酶广泛应用于众多行业。在食品加工中,果胶酶用于澄清果汁,蛋白酶用于嫩化肉类。在洗涤剂制造中,脂肪酶和纤维素酶能增强去污能力。在医学诊断中,葡萄糖氧化酶等酶被用于生物传感器,以测量血糖浓度。酶联免疫吸附试验(ELISA)依赖酶‑抗体结合物来检测特定抗原或抗体,例如在艾滋病病毒(HIV)检测中。
Drug design increasingly exploits enzyme inhibitors. For example, angiotensin‑converting enzyme (ACE) inhibitors treat hypertension, and reverse transcriptase inhibitors manage HIV infections. Understanding the kinetics and inhibition of enzymes is therefore not only academically important but also directly relevant to modern pharmacology and therapeutics.
药物设计越来越利用酶抑制剂。例如,血管紧张素转化酶(ACE)抑制剂用于治疗高血压,逆转录酶抑制剂用于控制艾滋病病毒感染。因此,理解酶的动力学和抑制作用不仅在学术上重要,而且与现代药理学和治疗学直接相关。
Published by TutorHao | Biology Revision Series | aleveler.com
更多咨询请联系16621398022(同微信)