Microbial Biodiesel Production via Lipase-Induced Autolysis

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Solution Overview

Problem

Current methods for producing biodiesel face challenges in scaling up production due to difficulties in obtaining large amounts of fatty acids and the long growth period of plants used for vegetable fatty acids, as well as complexities in metabolic engineering for enhanced production.

Innovation Solution

A method involving the culture of Rhodococcus opacus transformed with genes encoding triacyl glycerol lipase and monoacyl glycerol lipase to induce autolysis and convert oil into free fatty acids, which are then converted into fatty acid alkyl esters using an alcohol, enhancing productivity and efficiency.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If vegetable oils and fats are used to produce biodiesel through chemical processing, then the production process is well-established, but it is difficult to produce biodiesel in large amounts

Engineering Contradiction:
Improvebiodiesel production amountVSAvoidavailability of vegetable oils and fats
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The microorganism produces oil within its own cells and then uses its own lipase enzymes to hydrolyze this stored oil into free fatty acids, which are subsequently converted to biodiesel. This self-service mechanism eliminates the need for external vegetable oil sources and enables large-scale production through microbial cultivation.

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The microorganism acts as an intermediary system that converts readily available carbon sources into stored oil, then converts this oil into free fatty acids through autolysis, and finally transforms free fatty acids into biodiesel. This multi-stage biological conversion process overcomes the limitation of direct vegetable oil processing.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Quantity of substance

If oil is accumulated in microorganisms through carbon source enrichment, then oil production increases, but the growth period extends and other growth factors are depleted

Engineering Contradiction:
Improveoil accumulation in microorganismsVSAvoidmicrobial growth period
Core Design Contradiction:
Quantity of substanceVSDuration of action of moving object

Solution Approach 1:

The cultivation process is divided into distinct phases: a growth phase where microorganisms multiply and accumulate oil in their cells, followed by a harvest phase where the oil-containing biomass is collected and processed. This periodic approach allows optimization of both growth and oil accumulation without extending the overall production cycle indefinitely.

Inventive Principle:
Principle #19Periodic action

Solution Approach 2:

The microorganism is pre-engineered or pre-conditioned to possess high oil-producing capability and lipase activity before cultivation begins. This preliminary preparation ensures that during the growth phase, the microorganisms efficiently accumulate oil and can subsequently perform autolysis to release free fatty acids, reducing the need for extended processing time.

Inventive Principle:
Principle #10Preliminary action

3Productivity

If lipase genes are introduced into Rhodococcus opacus to enhance oil conversion, then free fatty acid production increases, but the genetic modification process becomes more complex

Engineering Contradiction:
Improvefree fatty acid production efficiencyVSAvoidgenetic modification process
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The introduced lipase genes enable the microorganism to perform multiple functions: storing oil in its cells during growth, hydrolyzing this stored oil into free fatty acids through autolysis, and providing the substrate for biodiesel production. This multi-functionality within a single organism simplifies the overall process despite the genetic modification complexity.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The oil storage function and oil hydrolysis function are merged within the same microorganism (Rhodococcus opacus). The microorganism both accumulates oil in its cells and contains lipase enzymes that hydrolyze this oil, eliminating the need for separate oil extraction and hydrolysis steps using different organisms or chemical processes.

Inventive Principle:
Principle #5Merging (Combining)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach allows for the high-efficiency production of fatty acid alkyl esters, which can be used as biodiesel, demonstrating improved productivity and environmental friendliness compared to existing methods.

Implementation Method 1

oil is an energy carrier that is synthesized and accumulated in microbial cells when microorganisms are rich in carbon sources but lack other growth factors (nitrogen, phosphorus, oxygen, sulfur, etc.)

Methodology Applied
Scientific EffectMetabolic accumulation: Fermentation

Implementation Method 2

inducing the autolysis of the produced oil in Rhodococcus opacus to produce a free fatty acid

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 3

autolysis of the produced oil in Rhodococcus opacus to produce a free fatty acid

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 4

converting the free fatty acid into an alkyl ester

Methodology Applied
Scientific EffectEsterification: Chemical Bonding

Data Source

PatentEP2455484B1Method for producing fatty acid alkyl esters using microorganisms having oil-producing ability
Publication Date: 2018.12.05 KOREA ADVANCED INST OF SCI & TECH
  • EP2455484B1 patent drawingFigure 1
  • EP2455484B1 patent drawingFigure 2
  • EP2455484B1 patent drawingFigure 3

AI summary

The present invention relates to a method of producing a fatty acid alkyl ester using microorganisms having the ability to produce oil, and more particularly to a method of producing a fatty acid alkyl ester, the method comprising culturing microorganisms having the ability to produce oil, thus accumulating a large amount of oil in the microorganisms, inducing the autolysis of the produced oil in the microorganisms to produce a free fatty acid, and converting the free fatty acid into an alkyl ester. According to the method of the present invention, oil accumulated in microorganisms, such as triacylglycerol that is typical oil produced by microorganisms, can be converted into a fatty acid alkyl ester with high efficiency using a metabolic engineering approach. Thus, the method of the present invention is useful for the industrial production of a fatty acid alkyl ester which has been recently found to be effective as biodiesel.