A method of preparing a variant lipolytic enzyme in the amino acid sequence in the host organism, encoded and having at least 90% sequence identity to a nucleotide sequence encoding a fungal lipolytic enzyme, the present invention, a Compared with the fungal lipolytic enzyme) groups in the amino acid sequence of at least the introduction of additional glycosylation sites) or one (one glycosylation site surface location b) on the polypeptide, and the active site of the enzyme ( Compared with the amino acid (at the original position of the outer loop of the catalytic triad distal to)) introduction of at least one amino acid is highly hydrophilic more c) or position 33,63,78,190, One modification, or to express a nucleotide sequence comprising a deletion in one or more of positions 307-319, or 311-312, at least at the position corresponding to the insertion or substitution at one or more of the 320 or 305 and 306 relates to a method that includes, in this case, correspond to the positions of the amino acid sequence of which when aligned with SEQ ID NO: 2, wherein said nucleotide sequence, the amino acid positions, fungi represented by SEQ ID NO: 23 or SEQ ID NO: 22 If it has at least 90% identity to the nucleotide sequence encoding the lipolytic enzyme, the modification is not a substitution at position 63, deletion is not in the position 311-312. Said nucleotide sequence has at least 90% identity with SEQ ID NO: 1, preferably. The present invention also relates to novel polypeptides and nucleic acids are produced by the method.本発明は、バリアント脂肪分解酵素を調製する方法であって、宿主生物において、真菌脂肪分解酵素をコードするヌクレオチド配列と少なくとも90%の配列同一性を有し、かつコードされたアミノ酸配列において、a)もとの真菌脂肪分解酵素と比較して、前記アミノ酸配列における少なくとも1つのグリコシル化部位(又は1つのさらなるグリコシル化部位)の導入;b)ポリペプチド上の表面位置、及び酵素の活性部位(触媒トリアッド)に対して遠位の外部ループ中の位置での(もとのアミノ酸と比較して)さらに親水性が高い少なくとも1つのアミノ酸の導入;又はc)位置33、63、78、190、305、306若しくは320の1又は2以上での置換若しくは挿入に相当する位置での少なくとも1つの修飾、又は位置311~312若しくは307~319の1又は2以上における欠失を含むヌクレオチド配列を発現させることを含む方法に関し、ここで、各アミノ酸位置は、配列番号2と整列させた場合のアミノ酸配列の位置に対応し、ここで、前記ヌクレオチド配列が、配列
