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CM-06

Functional analysis of the Phanerochaete chrysosporium Mnp gene promoter

and its application for apoptosis induction in Ganoderma lucidum

2
#,1
Y u -Hsuan Chen, Meng-Shuan Lai, Bang-Jau You* ,1

1 Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical
University, No. 91, Xueshi Rd., North Dist., Taichung City, Taiwan
2 National Chung Hsing University, No. 145, Xingda Rd., South Dist., Taichung City40227,
Taiwan

* E-mail: bangjau@yahoo.com.tw

Abstract
Ganoderma lucidum produces bioactive ganoderic acids, belonging to triterpenoid secondary
metabolites. Previous studies showed that aspirin and heat treatment induced apoptosis and
ganoderic acids production in G. lucidum. However, the role of apoptotic signaling in regulating
secondary metabolism remains unclear. We aimed to overexpress the pro-apoptotic protein Bax
in G. lucidum to investigate whether apoptosis signaling could induce ganoderic acid
production. Previous study indicated that constitutive Bax overexpression caused cell death
during gene transformation. Mnp (Manganese peroxidase) gene of P. chrysosporium was
induced by Mn²⁺ during nitrogen starvation. Promoter includes inverted CAAT motif, metal
response elements (MREs), heat shock elements (HREs), and activator protein 2 (AP-2) bind
sites. In this study, 1528bp Mnp promoter was cloned and recombinant DNA was constructed
to drive Bcl2 gene expression. Agrobacterium-mediated transformation was performed to
generate G. lucidum transformants. Promoter activity was evaluated under various culture
conditions. High expression of Bcl-2 was achieved in minimal liquid medium containing 180
μM or 900 μM Mn². These results confirmed that the P. chrysosporium Mnp promoter can be
induced by Mn²⁺ and nitrogen starvation conditions in G. lucidum. Application of this inducible
promoter to overexpress Bax will be conducted to investigate the role of apoptosis signaling on
secondary metabolites production.

Keywords: Phanerochaete chrysosporium; Inducible promoter; Mn²⁺ induction; Ganoderic
acids; Apoptosis