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After reading about how recombinant insulin is produced, the following question occured to me. Does the current level of technology allow any enzyme to be produced in a similar way?

As I see, producing amino acid sequences is not an issue. A possible difficulty I can think of is modifying the amino acid chains. In the case of insulin the problem has been solved. Are there some generally applicable solutions available to this problem, or do they have to be researched on a case-by-case basis?

Are there any other limitations to producing enzymes?

EDIT: As pointed out by P. Jay, the question might seem to suggests that insulin is an enzyme which it is not.

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    $\begingroup$ Anything can be produced, the problems lie in producing large amounts with high quality. $\endgroup$ – MattDMo Sep 13 '15 at 3:31
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No, not all enzymes (or other proteins for that matter) can be obtained in functional form by recombinant expression with today's methods. As you suspect, problems arise when complex post-translational modifications are necessary to obtain the correct function.

Direct modifications to peptide is one potential problem. Some of these can be resolved by expression the recombinant protein in the right cell type: for example, expression in a bacteria may not provide the right modification, but expression in yeast or a mammalian cell might. CHO cells are a common mammalian cell type used in such cases; antibody production in hybridomas is another well-known example. But there are plenty of posttranslational modifications that are poorly understood, and most likely there are still many completely unknown types; new ones are still being discovered.

Another posttranslational event is association with cell membranes. Membrane proteins (including membrane-embedded enzymes, for example in lipid metabolism) can be very difficult to express properly, as not only the peptide but some surround membrane structure must be reproduced in the expression system to obtain correct function. This is also a major hurdle in obtaining structures for membrane proteins, notably many cell surface receptors. This article is a summary of the current challenges and unsolved problems in this area. And you can imagine the problems with proteins that need to form large superstructures to be functional --- large protein polymers, respiratory chain complexes, ribosomes, DNA replication forks ...

So there is no universal method; all proteins are different, some are easy to express, some are extremely hard.

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If you are talking about the possibility, the answer is yes. However, you might have some difficulties in producing some proteins. In some cases, the reasons could be obvious--sugar modification and phosphorylation which can not happen in E. coli. Although phosphorylation is involved, if you do not know if your protein is phosphorylated, you would never find the reason why you can not get the active protein.

Polypeptide folds in the correct way or incorrect way in E Coli. Sometimes, it is difficult to fold proteins correctly in E coli expression systems. In addition, if your proteins has multiple polypeptide chains, it could be more difficult.

There are mammalian, yeast, insect expression systems, and these systems could be better if you have problems in producing proteins in E coli. But it is not always that these systems work.

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Yes enzymes can be produced and even be optimized like restriction enzymes which are now mostly recombinant thou to low efficiency or biosafety level of source bacteria leaving that behind ,the greatest problem here is the size of protein and it's gene as when you produce artificial bio products you've got a prokaryote host to do that in cases which they do it out of cells then it's completely different but for sure possible the greatest problem is efficiency of production....and by the way insulin is not an enzyme

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