Assembly of the LH2 Light-Harvesting Complexes of Thiorhodospira sibirica with Different Carotenoid Levels

摘要

Regardless of cultivation conditions, two types of LH2 complexes are always assembled in photosynthetic membranes of Thiorhodospira (T.) sibirica: short-wave B800-830 and long-wave B800-850. The present work studied the effect of qualitative and quantitative content of carotenoids on assembling of these complexes. Diphenylamine (DPA), an inhibitor of carotenoid biosynthesis, was used to decrease the content of carotenoids in photosynthetic membranes and the complexes. In the cells of T. sibirica strain A12(T), inhibition of carotenoid biosynthesis ranged from 40 to 50%. Complete suppression of carotenoid biosynthesis was achieved only once, which resulted in absorption spectrum changes of the DPA-LH2 complexes and lower stability of their structure; they were often destroyed during the isolation. The use of DPA with the cells of T. sibirica strain Kir-3 was more efficient. From inhibited cells of this strain, B800-850 and B800-830 complexes were isolated, in which the carotenoid content decreased to 80, 50, and 10%. Their spectral characteristics in the near infrared region were independent of the carotenoid quantity in the complexes and were similar to those of native complexes of the wild-type. At higher DPA concentration in the cultivation medium, accompanied by a decrease of the overall carotenoid content, such carotenoids of the earlier stages of biosynthesis as neurosporene, zeta-carotene, and their hydroxy derivatives, were found in the complexes. The efficiency of energy transfer from carotenoids to bacteriochlorophyll in all LH2 complexes was not shown to depend on the composition and quantity of carotenoids in the samples and was similar to 40%. Since the near-infrared spectrum of the LH2-complex of the B800-850 type was similar to that of the B800-850 complexes from Rhodospirillum molishianum, with 8 pairs of alpha/beta-heterodimers forming the basis of their structure, it is possible to assume that the structure of the studied B800-850 complex from T. sibirica also consists of 8 pairs of alpha/beta-heterodimers. A single carotenoid molecule is known to occur per the alpha/beta-heterodimer; therefore, on average less than one carotenoid molecule (8 x 0.1 = 0.8) occurs per an LH2 complex from T.sibirica with 10% carotenoid content. This implies that the population of LH2 complexes contains both carotenoidless LH2 complexes and the complexes containing one or more carotenoid molecules. Obviously, LH2 complexes of both types are assembled regardless of the quantity of carotenoids per a complex, and carotenoids are not the required component for their correct assembly in vivo.