藻胆蛋白生物合成研究进展
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摘要
藻胆蛋白作为藻类特殊的捕光色素蛋白复合体,有着独特的结构和功能.目前藻胆蛋白主要有2个来源:一是直接从天然藻体中分离提取;二是将合成藻胆蛋白的基因转到基因工程宿主中,通过体外重组方法获得.通过从天然藻体提取藻胆蛋白不但生产成本较高,而且副产物较多.通过体外重组来实现藻胆蛋白的生产不仅可以降低经济和时间成本,而且可以对藻胆蛋白的生物合成过程进行研究.此外,通过体外重组获得具有光学活性的藻胆蛋白时,可以添加不同的分子标签,从而拓展藻胆蛋白的应用范围.本文对藻胆蛋白的体内生物合成和体外重组研究的最新进展做了综述.
Photosynthesis has existed on the earth for about 4 billion years, and the photosynthetic organisms have emerged with the production of photosynthesis. In order to effectively capture light energy and adapt their living environment, algae and higher plants have evolved different light-harvesting antenna systems during the long evolution, one is the chlorophyll based light harvesting systems present mainly in terrestrial organisms, such as higher plants; and the other is the phycobiliprotein(PBP) based phycobilisome system mainly in aquatic organisms, such as cyanobacteria, red algae and cryptophytes. As a special light-harvesting pigment-protein complex, PBP can not only capture light energy, also shows many unique characteristics, such as good fluorescent properties and lots of pharmaceutical activities. Therefore, PBPs have been extensively studied and commercially used in foods, cosmetics, biotechnology detection, pharmacological and medicine fields. Due to its broad excitation spectrum, nontoxic, stable fluorescence and high quantum yield, PBPs have also employed as valuable fluorescent probes for immunological diagnosis. At present, there are mainly two ways to obtain fluorescent PBPs, one is to purify PBP directly from natural algae, and the other is to express recombinant PBP using genetic engineered host cells. Purification of PBPs from natural algae has many disadvantages, such as higher cost, complicated processes, and many by-products. Using genetically engineered cell biotechnology, large-scale and low-cost production of recombinant PBPs can be achieved, which can help solve problems such as source, quality control and so on. Since natural PBPs are mostly in the form of polymers such as trimers or hexamers, it is difficult to study the assembly process. Through genetic engineering techniques, monomers or subunits can be obtained, which will help to study the assembly process of PBP. In addition, novel recombinant PBPs can be produced by molecular design to improve their fluorescence and biological activity. The study of the biosynthesis will help to elucidate the relationship between the structure and spectral properties of PBPs and provide a basis for the construction of solar energy utilization systems. This review mainly focuses on the progress of combinational biosynthesis and in vivo reorganization of PBP in heterologous genetically engineered host cells, with emphasis on different genes coding for apo-PBPs and genes coding for different enzymes which catalyze the production of phycobilins from endogenous heme, along with the recent progress of three types of lyases which catalyze the covalently binding of phycobilins with apo-PBPs. The possible applications of these valuable recombinant PBPs were proposed as well.
Photosynthesis has existed on the earth for about 4 billion years, and the photosynthetic organisms have emerged with the production of photosynthesis. In order to effectively capture light energy and adapt their living environment, algae and higher plants have evolved different light-harvesting antenna systems during the long evolution, one is the chlorophyll based light harvesting systems present mainly in terrestrial organisms, such as higher plants; and the other is the phycobiliprotein(PBP) based phycobilisome system mainly in aquatic organisms, such as cyanobacteria, red algae and cryptophytes. As a special light-harvesting pigment-protein complex, PBP can not only capture light energy, also shows many unique characteristics, such as good fluorescent properties and lots of pharmaceutical activities. Therefore, PBPs have been extensively studied and commercially used in foods, cosmetics, biotechnology detection, pharmacological and medicine fields. Due to its broad excitation spectrum, nontoxic, stable fluorescence and high quantum yield, PBPs have also employed as valuable fluorescent probes for immunological diagnosis. At present, there are mainly two ways to obtain fluorescent PBPs, one is to purify PBP directly from natural algae, and the other is to express recombinant PBP using genetic engineered host cells. Purification of PBPs from natural algae has many disadvantages, such as higher cost, complicated processes, and many by-products. Using genetically engineered cell biotechnology, large-scale and low-cost production of recombinant PBPs can be achieved, which can help solve problems such as source, quality control and so on. Since natural PBPs are mostly in the form of polymers such as trimers or hexamers, it is difficult to study the assembly process. Through genetic engineering techniques, monomers or subunits can be obtained, which will help to study the assembly process of PBP. In addition, novel recombinant PBPs can be produced by molecular design to improve their fluorescence and biological activity. The study of the biosynthesis will help to elucidate the relationship between the structure and spectral properties of PBPs and provide a basis for the construction of solar energy utilization systems. This review mainly focuses on the progress of combinational biosynthesis and in vivo reorganization of PBP in heterologous genetically engineered host cells, with emphasis on different genes coding for apo-PBPs and genes coding for different enzymes which catalyze the production of phycobilins from endogenous heme, along with the recent progress of three types of lyases which catalyze the covalently binding of phycobilins with apo-PBPs. The possible applications of these valuable recombinant PBPs were proposed as well.
引文
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2 Adir N. Elucidation of the molecular structures of components of the phycobilisome:Reconstructing a giant. Photosynth Res, 2005, 85:15–32
3 Gantt E, Conti S F. The ultrastructure of porphyridium cruentum. J Cell Biol, 1965, 26:365
4 Gantt E, Conti S F. Phycobiliprotein localization in algae. Brookhaven Symp Biol, 1966, 19:393
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6 Li W J,PuY,Niu Z,etal.Structural andfunctionalinvestigation ofallophycocyanin trimer(in Chinese). BullSciTechnol, 2017, 62:1699–1713[李文军,蒲洋,牛壮,等.重组别藻蓝蛋白三聚体结构与功能.科学通报, 2017, 62:1699–1713] 7Zhang J, Ma J, Liu D, et al. Structure of phycobilisome from the red alga Griffithsia pacifica. Nature, 2017, 551:57
8 ChenY.Studiesoncyanobacteriochromesandthebiosynthesisofphycoerythrin(inChinese).DoctorDissertation.Wuhan:Huazhong UniversityofScienceandTechnology,2012[陈煜.蓝细菌光敏色素及藻红蛋白的生物合成研究.博士学位论文.武汉:华中科技大学, 2012]
9 Lundell D J, Glazer A N. Allophycocyanin B. A common beta subunit in Synechococcus allophycocyanin B(lambda max 670 nm)and allophycocyanin(lambda max 650 nm). J Biol Chem, 1981, 256:12600
10 Maccoll R. Cyanobacterial phycobilisomes. J Struct Biol, 1998, 124:311
11 Yi J J, Zang X N, Zhang X C, et al. Recombinant expression of a fluorescent phycocyanin holo-α-subunit from Arthrospira platensis in Escherichia coli(in Chinese). Period Ocean Univ China, 2011, 41:59–62[衣俊杰,臧晓南,张学成,等.具荧光活性的节旋藻藻蓝蛋白α亚基在大肠杆菌中的重组表达.中国海洋大学学报, 2011, 41:59–62]
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