Nowadays, it is not overly expensive to get a synthetic construct for a gene of interest, with your codons optimized for E.Coli expression. I got a huge improvement in the expression of a protein - like 20-50-fold better than when I was trying to express with E.Coli Rosetta (which in principle help for the expression of rare codons). Take a look at GeneScript or GeneArt (LifeTechnologies), prices are quite competitive.
Is it a soluble protein or a membrane protein? If you use a purification tag, which one is it and in which side of the protein? Is your protein full length or are you trying to express only a subpart of the protein?
And few clues:
Do you see a lot of your protein in the non soluble fraction in your SDS-Page? or no protein at all ? Synthetic gene or subcloning? Have you checked codon usage to avoid lots of "very rare" codons (sometimes usefull in an heterologus system)?
You can change the host system. IPTG is an inducer of BL21 DE3 strain. If possible, try GJ1158 strain. Its inducer is NaCl and media required for GJ 1158maintenance is saltfree media.
Actually...it very hard to express directly protein from eukaryotic protein into prokaryotic expression system...it is because in prokaryotic system dont have translation modification step...so...better you extract the total RNA from that fungal...do RT-PCR to obtain cDNA sequence...before clone and express into e.coli..
Totally agree with @Abdul Rahim, I recommend to synthesize DNA sequence and optimize codon suitable for E-coli usage. Then, try different strains of the host to find the best one. Good luck.
Nowadays, it is not overly expensive to get a synthetic construct for a gene of interest, with your codons optimized for E.Coli expression. I got a huge improvement in the expression of a protein - like 20-50-fold better than when I was trying to express with E.Coli Rosetta (which in principle help for the expression of rare codons). Take a look at GeneScript or GeneArt (LifeTechnologies), prices are quite competitive.