Overexpression of bacterial ethylene-forming enzyme gene in Trichoderma reesei enhanced the production of ethylene
2010
Xi Chen, Yong Liang, Jing Hua, Li Tao, Wensheng Qin, Sanfeng Chen
<p>In order to efficiently utilize natural cellulose materials to produce ethylene, three expression vectors containing the ethylene-forming enzyme (<i>efe</i>) gene from <i>Pseudomonas syringae</i> pv. <i>glycinea</i> were constructed. The target gene was respectively controlled by different promoters: <i>cbh</i> I promoter from<i> Trichoderma reesei </i>cellobiohydrolases I gene, <i>gpd</i> promoter from <i>Aspergillus nidulans</i> glyceraldehyde-3-phosphate dehydrogenase gene and <i>pgk</i> I promoter from <i>T. reesei </i>3-phosphoglycerate kinase I gene. After transforming into <i>T. reesei</i> QM9414, 43 stable transformants were obtained by PCR amplification and ethylene determination. Southern blot analysis of 14 transformants demonstrated that the <i>efe</i> gene was integrated into chromosomal DNA with copy numbers from 1 to 4. Reverse transcription polymerase chain reaction (RT-PCR) analysis of 6 transformants showed that the heterologous gene was transcribed. By using wheat straw as a carbon source, the ethylene production rates of aforementioned 14 transformants were measured. Transformant C30-3 with <i>pgk </i>I promoter had the highest ethylene production (4,012 nl h<sup>-1</sup> l<sup>-1</sup>). This indicates that agricultural wastes could be used to produce ethylene in recombinant filamentous fungus <i>T. reesei</i>.</p>
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