Improved production of human type II procollagen in the yeast Pichia pastoris in shake flasks by a wireless-controlled fed-batch system

Table 1 Review of OD₆₀₀, collagen II expression, C-P4H activities, and the amount of total fed methanol in four independent experiments with parallel predetermined constant feed. The pulse feeding method was used as a reference.

Exp.no.	OD₆₀₀ 46/72 h		Pepsin resistant collagen II		C-P4H/(dpm/50 μg protein)		Methanol feed/ml
	Feed	Reference	Feed	Reference	Feed	Reference	Feed	Reference
1	14.1	15.4	+	++	-	-	109	72
1A	12.2	15.4	+	++	-	-	>100	72
2	11.0/18.7	10.4/14.2	+++	n.d.	15449*	2042*	115	74
3	18.5	17.4	++++	++	13403	1526	85	75
4	19.7	14.6	++	+	6263	3391	104	89
4A	17.8		++		5238		102

Experiments 1, 2, 3, 4 and 4A have been performed with a quasi-continuous predetermined feeding rate, however in experiment 4A the methanol concentration was doubled compared to the other experiments. Experiment 1A was performed as a pO₂-stat (computer controlled methanol addition in dependence on the pO₂ signal). The amount of correctly folded collagen II (pepsin resistant) was evaluated from the SDS-PAGE gels. Highest values of C-P4H activity are shown as dpm per 50 μg of soluble protein. Total methanol feed in the cultivations is shown in the last column.
* 45 h

ISSN: 1472-6750