Method for separating and extracting pediocin PA-1 from fermentation solution

Abstract

The invention relates to a method for separating and extracting pediocin PA-1 from a fermentation solution. The method comprises the following steps: inactivating protease of the fermentation solution, releasing the pediocin PA-1 in the fermentation solution, performing plate-frame filtration to remove bacteria and granular impurities, performing microfiltration to remove high molecular weight impurities and performing two-step ultrafiltration: namely, the first step: selecting an ultrafiltration membrane with the molecular weight cutoff of 10000-30000 daltons with the main purpose of removing high molecular weight proteins, nucleic acid, colloid and the like; the second step: selecting the ultrafiltration membrane with the molecular weight cutoff of 1000-3000 daltons with the main purposes of removing small molecular weight impurities and concentrating. A concentrated solution is finally purified by C18 preparation chromatography and freeze-dried or added with common salt filler for spray-drying to get a powdery finished product. The process method disclosed by the invention has the advantages of whole-process fluid operation, low energy consumption, low labor intensity, mild conditions, easy industrial amplification, stable product quality, no consumption of an organic solvent, safety and environment-friendly property, and is suitable for a variety of follow-up purification methods.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and relates to a membrane separation technology, in particular to a method for separating and extracting pediocin lactate PA-1 from fermentation broth. Background technique [0002] Pediocin PA-1 is a small peptide with antibacterial activity consisting of 44 amino acids, with an isoelectric point of 9.6 and a theoretical molecular weight of 4624Da. Pediococcus lactis PA-1 was first discovered in Pediococcus lactis PAC1.0 and was named accordingly. With the deepening of research, it was found that in addition to Pediococcus lactis, other genera and species of lactic acid bacteria, such as Lactobacillus plantarum and Pediococcus parvulus, can also produce lactobacillus PA-1. [0003] The bacteriostatic scope of pediocin lactide PA-1 is lactic acid bacteria with close relatives and some Gram-positive bacteria that are not lactic acid bacteria, especially for common pathogenic bacteria and spoil...

AI Technical Summary

Problems solved by technology

[0004] At present, there are many reports on the extraction and purification methods of pediocin lactate PA-1. Similar to ordinary protein purification, most of them are the concentration of bacteriocin combined with chromatographic purification at various levels. These methods are suitable for small-scale purification in laboratories and can achieve ideal Purity, but the steps are cumbersome, the yield loss is large, and it is time-consuming and laborious

In 2008, Naghmouchi et al first reported the one-step purification of pediocin lactate PA-1 by affinity chromatography. This method is simple, fast, specific and reproducible, but the cost of affinity chromatography packing material is expensive and regeneration is difficult.

In any case, the existing method of salting out combined with chromatographic purification at various stages is too cumbersome, and the cost of a large amount of purification is too high; therefore, it is necessary to develop an industrial-grade purification method that is simpler and more practical

[0006] Membrane separation technology has the functions of separation, concentration, purification and refining. It is the most eye-catching high-tech in the era of circular economy. The emergence of membrane separation technology provides a new and feasible idea for industrial scale purification of bacteriocins. Separation and extraction of pediocin lactide PA-1 from fermentation broth by membrane separation technology has not been reported yet

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