Pregnenolone is a key intermediate in the biosynthesis of many steroid hormones and neuroprotective steroids. Sulfotransferase family cytosolic 2B member 1 (SULT2B1a) has been reported to be highly selective to sulfate pregnenolone. This study aimed to clarify the effect of missense single nucleotide polymorphisms (SNPs) of the human SULT2B1 gene on the sulfating activity of coded SULT2B1a allozymes toward Pregnenolone. To investigate the effects of single nucleotide polymorphisms of the SULT2B1 gene on the sulfation of pregnenolone by SULT2B1a allozymes, 13 recombinant SULT2B1a allozymes were generated, expressed, and purified using established procedures. Human SULT2B1a SNPs were identified by a comprehensive database search. 13 SULT2B1a nonsynonymous missense coding SNPs (cSNPs) were selected, and site-directed mutagenesis was used to generate the corresponding cDNAs, packaged in pGEX-2TK expression vector, encoding these 13 SULT2B1a allozymes, which were bacterially expressed in BL21 E. coli cells and purified by glutathione-Sepharose affinity chromatography. Purified SULT2B1a allozymes were analyzed for sulfating activities towards pregnenolone. In comparison with the wild-type SULT2B1a, of the 13 allozymes, 11 showed reduced activity toward pregnenolone at 0.1 µM. Specifically, P134L and R259Q allozymes, reported to be involved in autosomal-recessive congenital ichthyosis, displayed low activity (1–10%) toward pregnenolone. The findings of this study may demonstrate the impact of genetic polymorphism on the sulfation of pregnenolone in individuals with different SULT2B1 genotypes.

孕烯醇酮是许多类固醇激素和神经保护类固醇生物合成的关键中间体。据报道，磺基转移酶家族胞质 2B 成员 1 (SULT2B1a) 对硫酸盐孕烯醇酮具有高度选择性。本研究旨在阐明人类SULT2B1基因的错义单核苷酸多态性 (SNP) 对编码的 SULT2B1a 同种酶对孕烯醇酮的硫酸化活性的影响。为了研究SULT2B1基因的单核苷酸多态性对 SULT2B1a 同种酶硫酸化孕烯醇酮的影响，使用既定程序生成、表达和纯化了 13 种重组 SULT2B1a 同种酶。通过全面的数据库搜索鉴定了人类 SULT2B1a SNP。选择13个SULT2B1a非同义错义编码SNP（cSNP），采用定点诱变生成相应的cDNA，包装于pGEX-2TK表达载体中，编码这13个SULT2B1a同种酶，在BL21大肠杆菌细胞中细菌表达，通过谷胱甘肽-琼脂糖亲和层析纯化。分析纯化的 SULT2B1a 同种酶对孕烯醇酮的硫酸化活性。与野生型 SULT2B1a 相比，13 种同种酶中，有 11 种在 0.1 µM 浓度下对孕烯醇酮的活性降低。具体而言，据报道，P134L 和 R259Q 同种酶与常染色体隐性先天性鱼鳞病有关，对孕烯醇酮表现出低活性 (1-10%)。本研究的结果可能证明遗传多态性对不同SULT2B1基因型个体中孕烯醇酮硫酸化的影响。