Question

Lipase is a versatile industrial enzyme that can be applied to various industries. It is one of the most important classes of hydrolytic enzymes that catalyze the hydrolysis of triacylglycerol to diacylglycerol, fatty acids and glycerol. You are a bioprocess engineer assigned by your superior to work with a biotechnologist to provide a proposal on the upstream and downstream processing of microbial lipase targeted to the biodiesel processing application. To begin the project, you are required to present a preliminary investigation report on the following. a. Select a suitable microorganism and a lipid carbon source that can be applied to produce microbial lipase for the targeted application economically at industrial scale with justifications.

I have found a microorganism which can produce Microbial Lipase for biodiesel process application economically at industrial scale by doing research. I found out an article of microorganism name 'Streptomyces sp CS 326'.

The heading of the article says 'A neutral lipase applicable in biodiesel production from a newly isolated Streptomyces sp CS 326.'

what is meant by 'neutral lipase'? Is it different from Microbial lipase? And can i use this microorganism 'Streptomyces sp CS 326' for the production of Microbial lipase for biodiesel process application?

Answer 1

Characterization of Neutral Lipase BT-1 Isolated from the Labial Gland of Bombus terrestris Males

In addition to their general role in the hydrolysis of storage lipids, bumblebee lipases can participate in the biosynthesis of fatty acids that serve as precursors of pheromones used for sexual communication.

The study of temporal dynamics of lipolytic activity in crude extracts from the cephalic part of Bombus terrestris labial glands. Extracts from 3-day-old males displayed the highest lipolytic activity. The highest lipase gene expression level was observed in freshly emerged bumblebees, and both gene expression and lipase activity were lower in bumblebees older than 3 days. Lipase was purified from labial glands, further characterized and named as BT-1. The B. terrestris orthologue shares 88% sequence identity with B. impatiens lipase HA. The molecular weight of B. terrestris lipase BT-1 was approximately 30 kDa, the pH optimum was 8.3, and the temperature optimum was 50°C. Lipase BT-1 showed a notable preference for C8-C10 p-nitrophenyl esters, with the highest activity toward p-nitrophenyl caprylate (C8). The Michaelis constant (Km) and maximum reaction rate (Vmax) for p-nitrophenyl laurate hydrolysis were Km = 0.0011 mM and Vmax = 0.15 U/mg.

Also use in Biofuel production

In an attempt to isolate a biocatalyst able to catalyze biodiesel production from microbial source, Streptomyces sp. CS326 was screened from hundreds of soil isolates collected from various parts of Korea. In 16S rRNA sequence analysis, the strain showed high degree of similarity with Streptomyces xanthocidicus (99.79%); therefore, it is classified as Streptomyces sp. CS326. An extracellular lipase produced by the strain (LP326) was purified using a single step gel permeation chromatography on Sepharose CL-6B. Molecular weight of LP326 was estimated to be 17,000 Da by SDS-PAGE. The activity was optimum at 40 °C and pH 7.0 and was stable at pH 5.0-8.0 and below 50 °C. It preferred p-nitrophenyl palmitate (C16), a long chain substrate; and K (m) and V (max) for the substrate were determined to be 0.24 mM and 4.6 mM/min mg, respectively. First 10 N-terminal amino acid sequences were APDLVALQSE, which are different from so far reported lipases. LP326 catalyzed biodiesel production using methanol and various oils; therefore, the enzyme can be applicable in the field of biofuel.