Nakari-Setälä T, Aro N, Kalkkinen N, Alatalo E, Penttilä M
VTT Biotechnology and Food Research, Finland.
Eur J Biochem. 1996 Jan 15;235(1-2):248-55. doi: 10.1111/j.1432-1033.1996.00248.x.
The hfb1 gene of the filamentous fungus Trichoderma reesei, previously cloned as a gene which was abundantly expressed when the fungus was grown on glucose-containing medium, was shown to encode a novel fungal hydrophobin. The encoded 97-amino-acid protein is cysteine-rich and has a typical signal sequence for secretion. Signal-sequence cleavage and putative proteolytic processing results in the mature HFBI protein of 75 amino acids. Antibodies raised against the HFBI protein expressed in Escherichia coli detected the T. reesei HFBI protein in the fungal cell wall and in the culture medium of submerged glucose-containing cultures. The identity of HFBI was verified by N-terminal and peptide sequencing of proteins purified both from the cell wall and culture medium. In the cell wall most of the HFBI formed SDS-insoluble complexes that could be extracted with trifluoroacetic acid. Bubbling or freezing of the culture medium caused HFBI to form aggregates that coprecipitated with a yellow pigment produced by the fungus.
丝状真菌里氏木霉的hfb1基因，先前被克隆为该真菌在含葡萄糖培养基上生长时大量表达的一个基因，结果表明它编码一种新型真菌疏水蛋白。所编码的97个氨基酸的蛋白质富含半胱氨酸，并有一个典型的分泌信号序列。信号序列切割和假定的蛋白水解加工产生了75个氨基酸的成熟HFBI蛋白。针对在大肠杆菌中表达的HFBI蛋白产生的抗体，在真菌细胞壁和含葡萄糖深层培养的培养基中检测到了里氏木霉HFBI蛋白。通过对从细胞壁和培养基中纯化的蛋白质进行N端测序和肽段测序，验证了HFBI的身份。在细胞壁中，大部分HFBI形成了不溶于十二烷基硫酸钠（SDS）的复合物，可用三氟乙酸提取。培养基鼓泡或冷冻会导致HFBI形成聚集体，这些聚集体与真菌产生的一种黄色色素共沉淀。
