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体外生物转化（in vitro BioTransformation，ivBT）：生物制造的新前沿
石婷1,2,3, 宋展1,2,3,4, 宋世怡1,2,5, 张以恒1,2,3
- 1.中国科学院天津工业生物技术研究所低碳合成工程生物学重点实验室，天津 300308
2.中国科学院天津工业生物技术研究所体外合成生物学中心，天津 300308
3.合成生物学海河实验室，天津 300308
4.上海交通大学生命科学技术学院，微生物代谢国家重点实验室，上海 200240
5.华东理工大学生物反应器工程国家重点实验室，上海 200237
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收稿日期:2024-01-07修回日期:2024-03-11出版日期:2024-04-10 -
通讯作者:张以恒 -
作者简介:石婷 （1984—），女，博士，中国科学院天津工业生物技术研究所副研究员。2008年本科毕业于合肥工业大学生物与食品工程学院，2010年和2014年分别获得天津大学化工学院生物化工专业硕士和博士学位。主要研究方向为体外合成生物学、酶工程与微生物代谢工程。E-mail：shi_ting@tib.cas.cn宋展 （1996—），女，在读博士生。研究方向为体外合成生物学、酶工程和代谢工程。E-mail：song_zhan@sjtu.edu.cn张以恒 （1971—），男，博士，中国科学院天津工业生物技术研究所低碳合成工程生物学重点实验室主任，曾任美国弗吉尼亚理工大学终身正教授。1993年和1996年分别获得华东理工大学生物工程专业学士和硕士学位；2002年获得美国达特茅斯学院化学工程专业博士学位。主要研究方向为体外合成生物学、新质生物制造、生物炼制以及淀粉储能与释能。E-mail：zhang_xw@tib.cas.cn -
基金资助:科技部重点专项(2022YFA0912300);国家自然科学基金面上项目(NSFC32271544);合成生物学海河实验室颠覆性创新项目(22HHSWSS000155);天津市合成生物技术创新能力提升行动项目(TSBICIP-CXRC-067)
In vitro BioTransformation （ivBT）: new frontier of industrial biomanufacturing
Ting SHI1,2,3, Zhan SONG1,2,3,4, Shiyi SONG1,2,5, Yi-Heng P. Job ZHANG1,2,3
- 1.Key Laboratory of Engineering Biology for Low-Carbon Manufacturing，Tianjin Institute of Industrial Biotechnology，Chinese Academy of Sciences，Tianjin 300308，China
2.In Vitro Synthetic Biology Center，Tianjin Institute of Industrial Biotechnology，Chinese Academy of Sciences，Tianjin 300308，China
3.Haihe Laboratory of Synthetic Biology，Tianjin 300308，China
4.State Key Laboratory of Microbial Metabolism，School of Life Sciences and Biotechnology，Shanghai JiaoTong University，Shanghai 200240，China
5.State Key Laboratory of Bioreactor Engineering，East China University of Science and Technology，Shanghai 200237，China
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Received:2024-01-07Revised:2024-03-11Online:2024-04-10 -
Contact:Yi-Heng P. Job ZHANG
摘要：
人类社会的重大挑战（如粮食安全、能源安全、气候变化与双碳目标等）驱动全社会寻求创新型技术解决方案。体外生物转化（in vitro BioTransformation，ivBT）是介于微生物发酵与酶催化之间的新质生物制造平台，多酶分子机器是其超限生物催化剂。它基于大道至简原则，利用多个天然酶、人工酶以及（仿生/天然）辅酶等重构生化途径，摆脱生物体生存局限（如细胞复制、基础代谢、复杂调控和能量供给等），超越细胞合成极限，实现重要生物转化与超限能量转换，尤其是生产低值大宗产品与新能源产品等。工业生物制造的三个平台技术分别是基于细胞工厂的发酵、基于酶分子的生物催化与基于多酶分子机器的ivBT。本综述对ivBT给出明确定义，阐明其多酶途径设计原则与产业化技术研发路径，比较该平台与现有生物制造平台相似性与不同点，介绍多个代表性案例，以及讨论其未来的机会与挑战。ivBT技术发展采用设计-构建-判决-优化的线性策略，开发能够满足国家需求的超高效多酶分子机器。利用ivBT有望形成超过30万亿元生物产品的工业生物制造，助力实现人类社会的多项重要需求，如粮食安全、新型能源体系等。人造淀粉不仅将帮助中国端牢粮食饭碗，而且将是一个全新且安全的高密度储氢载体（比压缩氢气高2.5倍）与高能储电介质（比锂电池高10倍）。
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引用本文
石婷, 宋展, 宋世怡, 张以恒. 体外生物转化（in vitro BioTransformation，ivBT）：生物制造的新前沿[J]. 合成生物学, DOI: 10.12211/2096-8280.2024-004.
Ting SHI, Zhan SONG, Shiyi SONG, Yi-Heng P. Job ZHANG. In vitro BioTransformation （ivBT）: new frontier of industrial biomanufacturing[J]. Synthetic Biology Journal, DOI: 10.12211/2096-8280.2024-004.
图1 生物制造平台分类
Fig. 1 Classification for industrial biomanufacturing platforms
图1 生物制造平台分类
Fig. 1 Classification for industrial biomanufacturing platforms
表1 生物制造平台的比较
Table 1 Comparison of biomanufacturing platforms
| 生物制造平台 | 微生物发酵 （Fermentation） | 酶催化 （Biocatalysis） | 体外生物转化 （ivBT) |
|---|---|---|---|
| 催化剂 | 细胞工厂 | 酶分子、级联多酶 | 多酶分子机器 |
| 标志性产品 | 初级代谢产物 次级代谢产物 生物大分子 微生物蛋白 | 果葡糖浆 生物质糖 医药原料 NMN | 肌醇、塔格糖 合成淀粉 糖水制绿氢 糖酶燃料电池 |
| 产品种类 | 极多 | 较多 | 较少 |
| 优选水解、异构、手性合成等方式合成产品 | 优选异构、糖苷键重排、分解与合成代谢等方式合成产品 | ||
| 产品规模 | 小、中、大 | 小 | 超大（如能源、粮食） |
| 浓度（Titer） | 低 | 高 | 高 |
| 得率（Yield） | 较低 | 高 | 高 |
| 速率（Rate） | 低 | 最高 | 高 |
| 生物安全 | 有挑战、强监管 | 好 | 最好 |
| 技术壁垒 | 低 | 高 | 最高 |
表1 生物制造平台的比较
Table 1 Comparison of biomanufacturing platforms
| 生物制造平台 | 微生物发酵 （Fermentation） | 酶催化 （Biocatalysis） | 体外生物转化 （ivBT) |
|---|---|---|---|
| 催化剂 | 细胞工厂 | 酶分子、级联多酶 | 多酶分子机器 |
| 标志性产品 | 初级代谢产物 次级代谢产物 生物大分子 微生物蛋白 | 果葡糖浆 生物质糖 医药原料 NMN | 肌醇、塔格糖 合成淀粉 糖水制绿氢 糖酶燃料电池 |
| 产品种类 | 极多 | 较多 | 较少 |
| 优选水解、异构、手性合成等方式合成产品 | 优选异构、糖苷键重排、分解与合成代谢等方式合成产品 | ||
| 产品规模 | 小、中、大 | 小 | 超大（如能源、粮食） |
| 浓度（Titer） | 低 | 高 | 高 |
| 得率（Yield） | 较低 | 高 | 高 |
| 速率（Rate） | 低 | 最高 | 高 |
| 生物安全 | 有挑战、强监管 | 好 | 最好 |
| 技术壁垒 | 低 | 高 | 最高 |
图2 ivBT 的设计原则
Fig. 2 Design principles of ivBT
图2 ivBT 的设计原则
Fig. 2 Design principles of ivBT
图3 ivBT 和 CEB 的生物制造产品规模
Fig. 3 Target product size for biomanufacturing of ivBT and CEB
图3 ivBT 和 CEB 的生物制造产品规模
Fig. 3 Target product size for biomanufacturing of ivBT and CEB
表2 ivBT 与相似技术的区分
Table 2 Comparison of ivBT with similar biotechnologies
| 体外生物转化（ivBT） | 多酶级联催化（CEB） | 无细胞蛋白质合成（CFPS） | |
|---|---|---|---|
| 目标 | 大规模生物制造 产品规模：>1万吨，甚至10亿吨级 | 精细生物制造 精细产品：～1000公斤级， <100吨 | 研究工具 特殊制造（快速生产克级蛋白质） |
| 代表性产品 | 粮食、能源、材料 （如淀粉、绿氢、肌醇、塔格糖） | 药物中间体（NMN） | 疫苗合成 |
| 产品市场规模 （每个产品） | 5亿（最小市场） →100亿（塔格糖） →10万亿（绿氢） | 千万（最大市场低于5亿） | NA |
| 目标产品数目 | ～100（粮食、能源等大宗产品） | ～10,000（精细化学品） | NA |
| 原料成本/ 产品价格 | >50%→90%（最大） | 5%→20% | NA |
| 合成途径设计 | 非天然途径与人造电子传递链 | 主反应与辅酶再生，利用部分天然途径 | 利用天然合成途径 |
| 催化元件 | 天然（超稳）酶、人工酶、固定化多酶、（仿生）辅酶再生 | 天然（常温）酶、改造酶、固定化酶、辅酶再生 | 细胞裂解液或纯化元件、外加氨基酸、ATP供体、DNA模板 |
| 元件需求 | 超低成本酶、超稳定固定化酶、价廉且稳定（仿生）辅酶 | 酶成本不敏感，利用天然辅酶 | 利用细胞裂解液中的有效成分 |
| 生产周期 | 天、周、月（多次，连续） | 小时、天（一次，极少多次） | 小时（一次） |
表2 ivBT 与相似技术的区分
Table 2 Comparison of ivBT with similar biotechnologies
| 体外生物转化（ivBT） | 多酶级联催化（CEB） | 无细胞蛋白质合成（CFPS） | |
|---|---|---|---|
| 目标 | 大规模生物制造 产品规模：>1万吨，甚至10亿吨级 | 精细生物制造 精细产品：～1000公斤级， <100吨 | 研究工具 特殊制造（快速生产克级蛋白质） |
| 代表性产品 | 粮食、能源、材料 （如淀粉、绿氢、肌醇、塔格糖） | 药物中间体（NMN） | 疫苗合成 |
| 产品市场规模 （每个产品） | 5亿（最小市场） →100亿（塔格糖） →10万亿（绿氢） | 千万（最大市场低于5亿） | NA |
| 目标产品数目 | ～100（粮食、能源等大宗产品） | ～10,000（精细化学品） | NA |
| 原料成本/ 产品价格 | >50%→90%（最大） | 5%→20% | NA |
| 合成途径设计 | 非天然途径与人造电子传递链 | 主反应与辅酶再生，利用部分天然途径 | 利用天然合成途径 |
| 催化元件 | 天然（超稳）酶、人工酶、固定化多酶、（仿生）辅酶再生 | 天然（常温）酶、改造酶、固定化酶、辅酶再生 | 细胞裂解液或纯化元件、外加氨基酸、ATP供体、DNA模板 |
| 元件需求 | 超低成本酶、超稳定固定化酶、价廉且稳定（仿生）辅酶 | 酶成本不敏感，利用天然辅酶 | 利用细胞裂解液中的有效成分 |
| 生产周期 | 天、周、月（多次，连续） | 小时、天（一次，极少多次） | 小时（一次） |
图4 ivBT 的体外肌醇合成途径（A） αGP—α-葡聚糖磷酸化酶；PGM—葡萄糖 6-磷酸异构酶；IPS—肌醇 3-磷酸合成酶；IMP—肌醇单磷酸磷酸酶； IA—异淀粉酶；（B）首个 ivBT 肌醇工业化生产工厂图片
Fig. 4 Inositol synthesis pathway of ivBT(A) αGP—α-glucan phosphorylase; PGM—phosphoglucomutase; IPS—inositol 3-phosphate synthase; IMP—inositol monophosphatase; IA—isoamylase; (B)Image of the first large-scale inositol factory
图4 ivBT 的体外肌醇合成途径（A） αGP—α-葡聚糖磷酸化酶；PGM—葡萄糖 6-磷酸异构酶；IPS—肌醇 3-磷酸合成酶；IMP—肌醇单磷酸磷酸酶； IA—异淀粉酶；（B）首个 ivBT 肌醇工业化生产工厂图片
Fig. 4 Inositol synthesis pathway of ivBT(A) αGP—α-glucan phosphorylase; PGM—phosphoglucomutase; IPS—inositol 3-phosphate synthase; IMP—inositol monophosphatase; IA—isoamylase; (B)Image of the first large-scale inositol factory
图5 ivBT 利用淀粉合成健康糖的人工合成途径αGP、PGM、IPS、IMP 同肌醇合成途径。PGI—葡萄糖 6-磷酸异构酶；FPP—果糖 6-磷酸磷酸酶；TPE—塔格糖 6-磷酸 4-差向异构酶；TPP—塔格糖 6-磷酸磷酸酶；MPI—甘露糖 6-磷酸异构酶；MPP—甘露糖 6-磷酸磷酸酶；API—阿洛酮糖 6-磷酸异构酶；APP—阿洛酮糖 6-磷酸磷酸酶
Fig. 5 Rare sugars artificial synthesis pathway of ivBTEnzymes of αGP, PGM, IPS, IMP are the same as inositol synthesis pathway. PGI—phosphoglucose isomerase; FPP—fructose 6-phosphatase; TPE—tagatose 6-phosphate 4-epimerase; TPP—tagatose 6-phosphatase; MPI—mannose 6-phosphate isomerase; MPP—mannose 6-phosphatase; API—allulose 6-phosphate isomerase; APP—allulose 6-phosphatase
图5 ivBT 利用淀粉合成健康糖的人工合成途径αGP、PGM、IPS、IMP 同肌醇合成途径。PGI—葡萄糖 6-磷酸异构酶；FPP—果糖 6-磷酸磷酸酶；TPE—塔格糖 6-磷酸 4-差向异构酶；TPP—塔格糖 6-磷酸磷酸酶；MPI—甘露糖 6-磷酸异构酶；MPP—甘露糖 6-磷酸磷酸酶；API—阿洛酮糖 6-磷酸异构酶；APP—阿洛酮糖 6-磷酸磷酸酶
Fig. 5 Rare sugars artificial synthesis pathway of ivBTEnzymes of αGP, PGM, IPS, IMP are the same as inositol synthesis pathway. PGI—phosphoglucose isomerase; FPP—fructose 6-phosphatase; TPE—tagatose 6-phosphate 4-epimerase; TPP—tagatose 6-phosphatase; MPI—mannose 6-phosphate isomerase; MPP—mannose 6-phosphatase; API—allulose 6-phosphate isomerase; APP—allulose 6-phosphatase
图 6 ivBT 的体外纤维素合成淀粉途径EG—内切葡聚糖酶；CBH—纤维二糖水解酶；CBP—纤维二糖磷酸化酶；PGP—马铃薯 α-葡聚糖磷酸化酶
Fig. 6 Cellulose-amylose synthesis pathway of ivBTEG—endoglucanase; CBH—cellobiohydrolase; CBP—cellobiose phosphorylase; PGP—potato α-glucan phosphorylase
图 6 ivBT 的体外纤维素合成淀粉途径EG—内切葡聚糖酶；CBH—纤维二糖水解酶；CBP—纤维二糖磷酸化酶；PGP—马铃薯 α-葡聚糖磷酸化酶
Fig. 6 Cellulose-amylose synthesis pathway of ivBTEG—endoglucanase; CBH—cellobiohydrolase; CBP—cellobiose phosphorylase; PGP—potato α-glucan phosphorylase
图7 ivBT 的体外 CO2 合成淀粉途径［96］AOX—醇氧化酶；FIS—甲醛酶；DAK—二羟基丙酮激酶；TIM—磷酸甘油醛异构酶；ALD—果糖 1，6-二磷酸醛缩酶；FBP—果糖 1，6-二磷酸酶；PGI—葡萄糖 6-磷酸异构酶；PGM—葡萄糖6-磷酸变位酶；AGP—ADP-葡萄糖焦磷酸化酶；SS—淀粉合成酶；CAT—过氧化氢酶；PPK—多聚磷酸激酶；PPA—焦磷酸酶
Fig. 7 CO2-Starch synthesis pathway of ivBT[96]AOX—alcohol oxidase; FIS—formolase; DAK—dihydroxyacetone kinase; TIM—triose phosphate isomerase; ALD—fructose-bisphosphate aldolase; FBP—fructose bisphosphatase; PGI—phosphoglucose isomerase; PGM—phosphoglucomutase; AGP—ADP-glucose pyrophosphorylase; SS—starch synthase; CAT—catalase; PPK—polyphosphate kinase; PPA—pyrophosphatase
图7 ivBT 的体外 CO2 合成淀粉途径［96］AOX—醇氧化酶；FIS—甲醛酶；DAK—二羟基丙酮激酶；TIM—磷酸甘油醛异构酶；ALD—果糖 1，6-二磷酸醛缩酶；FBP—果糖 1，6-二磷酸酶；PGI—葡萄糖 6-磷酸异构酶；PGM—葡萄糖6-磷酸变位酶；AGP—ADP-葡萄糖焦磷酸化酶；SS—淀粉合成酶；CAT—过氧化氢酶；PPK—多聚磷酸激酶；PPA—焦磷酸酶
Fig. 7 CO2-Starch synthesis pathway of ivBT[96]AOX—alcohol oxidase; FIS—formolase; DAK—dihydroxyacetone kinase; TIM—triose phosphate isomerase; ALD—fructose-bisphosphate aldolase; FBP—fructose bisphosphatase; PGI—phosphoglucose isomerase; PGM—phosphoglucomutase; AGP—ADP-glucose pyrophosphorylase; SS—starch synthase; CAT—catalase; PPK—polyphosphate kinase; PPA—pyrophosphatase
图8 淀粉体外合成氢气途径αGP 和 PGM 同肌醇合成途径。G6PDH—葡萄糖 6-磷酸脱氢酶；6PGL—6-磷酸葡萄糖内酯酶；6PGDH—6-磷酸葡萄糖酸脱氢酶；RPI—核糖 5-磷酸异构酶；RPE—核酮糖 5-磷酸 3-差向异构酶；TK—转酮酶；TAL—转醛酶；TIM—磷酸甘油醛异构酶；ALD—果糖1，6-二磷酸醛缩酶；FBP—果糖1，6-二磷酸酶；PGI—葡萄糖6-磷酸异构酶；SHI—氢酶；NROR—NADPH-红氧还蛋白氧化还原酶；DI—心肌黄酶。代谢物：g1p—葡萄糖 1-磷酸；g6p—葡萄糖 6-磷酸；ru5p—核酮糖 5-磷酸；x5p—木糖 5-磷酸；r5p—核糖 5-磷酸；s7p—景天庚酮糖 7-磷酸；g3p—甘油醛 3-磷酸；e4p—赤藓糖 4-磷酸；dhap—磷酸二羟丙酮；fdp—果糖1，6-二磷酸；f6p—果糖 6-磷酸。
Fig. 8 Hydrogen synthesis pathway of ivBTEnzymes of αGP and PGM are the same as inositol synthesis pathway. G6PDH—glucose 6-phosphate dehydrogenase; 6PGL—6-phosphogluconolactonase; 6PGDH—6-phosphogluconate dehydrogenase; RPI—ribose 5-phosphate isomerase; RPE—ribulose 5-phosphate 3-epimerase; TK—transketolase; TAL—transaldolase; TIM—triose phosphate isomerase; ALD—fructose-bisphosphate aldolase; FBP—fructosebisphosphatase; PGI—phosphoglucose isomerase; SHI—soluble hydrogenase I; NROR—NADPH rubredoxin oxidoreductase; DI—diaphorase. The metabolites are: g1p—glucose 1-phosphate; g6p—glucose 6-phosphate; ru5p—ribulose 5-phosphate; x5p—xylulose 5-phosphate; r5p—ribose 5-phosphate; s7p—sedoheptulose 7-phosphate; g3p—glyceraldehyde 3-phosphate; e4p—erythrose 4-phosphate; dhap—dihydroxacetone phosphate; fdp—fructose 1,6-diphosphate; f6p—fructose 6-phosphate.
图8 淀粉体外合成氢气途径αGP 和 PGM 同肌醇合成途径。G6PDH—葡萄糖 6-磷酸脱氢酶；6PGL—6-磷酸葡萄糖内酯酶；6PGDH—6-磷酸葡萄糖酸脱氢酶；RPI—核糖 5-磷酸异构酶；RPE—核酮糖 5-磷酸 3-差向异构酶；TK—转酮酶；TAL—转醛酶；TIM—磷酸甘油醛异构酶；ALD—果糖1，6-二磷酸醛缩酶；FBP—果糖1，6-二磷酸酶；PGI—葡萄糖6-磷酸异构酶；SHI—氢酶；NROR—NADPH-红氧还蛋白氧化还原酶；DI—心肌黄酶。代谢物：g1p—葡萄糖 1-磷酸；g6p—葡萄糖 6-磷酸；ru5p—核酮糖 5-磷酸；x5p—木糖 5-磷酸；r5p—核糖 5-磷酸；s7p—景天庚酮糖 7-磷酸；g3p—甘油醛 3-磷酸；e4p—赤藓糖 4-磷酸；dhap—磷酸二羟丙酮；fdp—果糖1，6-二磷酸；f6p—果糖 6-磷酸。
Fig. 8 Hydrogen synthesis pathway of ivBTEnzymes of αGP and PGM are the same as inositol synthesis pathway. G6PDH—glucose 6-phosphate dehydrogenase; 6PGL—6-phosphogluconolactonase; 6PGDH—6-phosphogluconate dehydrogenase; RPI—ribose 5-phosphate isomerase; RPE—ribulose 5-phosphate 3-epimerase; TK—transketolase; TAL—transaldolase; TIM—triose phosphate isomerase; ALD—fructose-bisphosphate aldolase; FBP—fructosebisphosphatase; PGI—phosphoglucose isomerase; SHI—soluble hydrogenase I; NROR—NADPH rubredoxin oxidoreductase; DI—diaphorase. The metabolites are: g1p—glucose 1-phosphate; g6p—glucose 6-phosphate; ru5p—ribulose 5-phosphate; x5p—xylulose 5-phosphate; r5p—ribose 5-phosphate; s7p—sedoheptulose 7-phosphate; g3p—glyceraldehyde 3-phosphate; e4p—erythrose 4-phosphate; dhap—dihydroxacetone phosphate; fdp—fructose 1,6-diphosphate; f6p—fructose 6-phosphate.
图 9 生物学研发模式：黑箱-灰箱-白箱
Fig. 9 The development of scientific research and development model: black-box, gray-box and white-box
图 9 生物学研发模式：黑箱-灰箱-白箱
Fig. 9 The development of scientific research and development model: black-box, gray-box and white-box
表3 ivBT 作为工业生物制造平台的技术优势
Table 3 Advantages of ivBT for biomanufacturing
| 细胞工厂的局限 | 体外生物转化的优势 | |
|---|---|---|
| 生长偶联 | 细胞生长繁殖与产品制造的耦合，存在有限资源竞争、分配与调控等难题 | 催化剂合成与产品制造时空分离，产品制造是唯一目标 |
| 产品得率 | 产品得率低（由于细胞繁殖、副产物生成等问题） | 近理论得率 |
| 能量限制 | 生物能量学限制，需 ATP与还原力的净合成 | ATP 与还原力的平衡，不浪费生物能量 |
| 反应速率 | 反应速度（Rxn）低 | Rxn提升10倍以上 |
生物大分子 合成 | 生物大分子不能通过细胞 | 没有细胞膜，大分子降解与合成耦合 |
特殊极性分 子合成 | 特殊极性分子合成如磷酸糖不能过细胞膜 | 没有细胞膜，自由扩散 |
| 三传限制 | 细胞体内三传“动量传递、热量传递、质量传递”限制 | 超限制造（微通道反应器）超越传统“三传”限制 |
表3 ivBT 作为工业生物制造平台的技术优势
Table 3 Advantages of ivBT for biomanufacturing
| 细胞工厂的局限 | 体外生物转化的优势 | |
|---|---|---|
| 生长偶联 | 细胞生长繁殖与产品制造的耦合，存在有限资源竞争、分配与调控等难题 | 催化剂合成与产品制造时空分离，产品制造是唯一目标 |
| 产品得率 | 产品得率低（由于细胞繁殖、副产物生成等问题） | 近理论得率 |
| 能量限制 | 生物能量学限制，需 ATP与还原力的净合成 | ATP 与还原力的平衡，不浪费生物能量 |
| 反应速率 | 反应速度（Rxn）低 | Rxn提升10倍以上 |
生物大分子 合成 | 生物大分子不能通过细胞 | 没有细胞膜，大分子降解与合成耦合 |
特殊极性分 子合成 | 特殊极性分子合成如磷酸糖不能过细胞膜 | 没有细胞膜，自由扩散 |
| 三传限制 | 细胞体内三传“动量传递、热量传递、质量传递”限制 | 超限制造（微通道反应器）超越传统“三传”限制 |
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