Table of Contents
What is lignin?
- Lignin is a class of complex organic polymers that constitute essential structural components in the connective tissues of most plants.
- Lignins are crucial to the construction of cell walls, notably in wood and bark, due to their ability to impart stiffness and resist decay. Chemically, lignins are polymers derived from phenolic precursors through cross-linking.
- Lignin was originally mentioned in 1813 by the Swiss botanist A. P. de Candolle, who characterized it as a tasteless, fibrous substance that is insoluble in water and alcohol but soluble in mild alkaline solutions and can be precipitated from solution with acid.
- He named the substance lignine, which is derived from the Latin word for wood, lignum. It is second only to cellulose in terms of the abundance of organic polymers on Earth.
- Lignin comprises 30 percent of non-fossil organic carbon on Earth and 20 to 35 percent of wood’s dry bulk.
- Lignin is detected in red algae, suggesting that lignin was also synthesized by the progenitor of plants and red algae. This study also shows that the initial purpose of lignin was structural, as it performs this role in the red alga Calliarthron, where it supports the joints between calcified segment joints.
Structure of lignin
- Lignin is a group of highly heterogeneous polymers produced from a small number of lignol precursors.
- The diversity and degree of crosslinking between these lignols give rise to heterogeneity. Coniferyl alcohol (4-hydroxy-3-methoxyphenylpropane) (G, its radical is sometimes referred to as guaiacyl), sinapyl alcohol (3,5-dimethoxy-4-hydroxyphenylpropane) (S, its radical is sometimes referred to as syringyl), and paracoumaryl alcohol (4-hydroxyphenylpropane) are the three main types of cross-linking lignols (H, its radical is sometimes called 4-hydroxyphenyl).
- The ratio of precursor “monomers” (lignols) varies depending on the plant source.
- Typically, lignins are categorized based on their syringyl/guaiacyl ratio. Lignin in gymnosperms (softwoods, grasses) is generated from coniferyl alcohol, which, following pyrolysis, produces guaiacol.
- Some of the coniferyl alcohol in angiosperms (hardwoods) is converted to sinapyl alcohol. Hence, lignin in angiosperms has both guaiacyl and syringyl constituents.
- The molecular masses of lignin exceed 10,000 u. It is hydrophobic because aromatic subunits predominate. Since the material is heterogeneous, it is difficult to measure the degree of polymerisation. According on the method of isolation, various kinds of lignin have been described.
- Several grasses are predominantly G, while certain palms are predominantly S.
- All lignins contain minute amounts of unfinished or modified monolignols, while non-woody plants are rich in other monomers.
Biological function of lignin
- Lignin fills the crevices between cellulose, hemicellulose, and pectin in the cell wall, particularly in vascular and support tissues: xylem tracheids, vessel elements, and sclereremic cells.
- Lignin is essential for water and aqueous nutrient transport in plant stems. The polysaccharide components of plant cell walls are hydrophilic and therefore water-permeable, whereas lignin is hydrophobic.
- Lignin’s crosslinking of polysaccharides prevents the cell wall from absorbing water. Consequently, lignin enables the vascular tissue of a plant to conduct water efficiently.
- Lignin is present in all vascular plants but absent in bryophytes, supporting the notion that lignin’s original role was limited to water transport.
- It is covalently bonded to hemicellulose and so cross-links various plant polysaccharides, thereby imparting mechanical strength to the cell wall and, by extension, the entire plant.
- Wood (mostly made of xylem cells and lignified sclerenchyma fibres) in vascular plants is held together by lignin. This is its most frequently cited function.
- Lastly, lignin confers disease resistance by accumulating at the site of pathogen infiltration, hence reducing the plant cell’s susceptibility to cell wall breakdown.
Factors affecting lignin degradation
The degradation of lignin is influenced by a variety of factors. Some of the important factors that affect lignin degradation are:
- Microbial diversity: The diversity and abundance of microorganisms present in the environment can have a significant impact on the rate and extent of lignin degradation. A high diversity of lignin-degrading microorganisms can result in more efficient degradation of lignin.
- Oxygen availability: The availability of oxygen can affect the rate of lignin degradation as many lignin-degrading microorganisms require oxygen for their metabolism. In anaerobic conditions, the degradation of lignin is slower.
- pH: The pH of the environment can also affect the rate of lignin degradation. Many lignin-degrading microorganisms have an optimal pH range for their activity, and changes in pH can affect their growth and activity.
- Temperature: Temperature is an important factor in lignin degradation as it affects the activity of lignin-degrading enzymes. Higher temperatures generally increase the rate of lignin degradation, but extreme temperatures can also denature enzymes and inhibit lignin degradation.
- Nutrient availability: The availability of nutrients such as carbon, nitrogen, and phosphorus can also influence the rate of lignin degradation. Many lignin-degrading microorganisms require these nutrients for their growth and metabolism.
- Lignin structure and complexity: The structure and complexity of the lignin molecule can also affect its degradation. Lignin that is more complex and has more cross-linkages between lignin molecules is generally more difficult to degrade.
- Moisture content: Moisture content can play an important role in lignin degradation, as it affects the availability of water for lignin-degrading microorganisms. Too little moisture can inhibit microbial activity, while too much moisture can create anaerobic conditions that may slow down lignin degradation.
- Added Nitrogen: Nitrogen is an essential nutrient for many lignin-degrading microorganisms, and adding nitrogen to the environment can increase the rate of lignin degradation. Nitrogen can also stimulate the growth of bacteria that support the growth of lignin-degrading fungi.
- Added Glucose: Adding glucose to the environment can also increase the rate of lignin degradation, as glucose is a readily available source of energy for lignin-degrading microorganisms. However, excessive glucose can lead to the accumulation of organic acids that can inhibit microbial activity.
- Aeration: Aeration is important for the growth of aerobic microorganisms that are involved in lignin degradation. Proper aeration can maintain the availability of oxygen, which is necessary for the activity of many lignin-degrading enzymes. Lack of aeration can result in anaerobic conditions that may slow down lignin degradation.
The biological breakdown of lignin is an extremely crucial component of the intricate carbon and oxygen cycle that occurs in our biosphere. Despite the fact that most of the lignin that plants produce is converted into CO2 and released back into the atmosphere, the microbiology behind the process of lignin degradation is still not comprehensively understood.
Interestingly, it has been discovered that a diverse range of microorganisms, including both fungi and bacteria, are capable of breaking down lignin. In addition to these, certain microorganisms such as cyanobacteria and actinomycetes are also known to play a significant role in lignin degradation, although the extent of their involvement varies significantly.
The degradation and transformation of lignocellulosic waste is largely dependent on the metabolism of native microorganisms, and different microbial populations dominate at different stages of the process, each with distinct roles to play in the degradation of organic matter.
1. Lignin-degrading bacteria
Lignin-degrading bacteria are a diverse group of microorganisms that have the ability to degrade lignin using a variety of enzymatic and non-enzymatic mechanisms. Some examples of lignin-degrading bacteria are:
- Streptomyces: These are aerobic bacteria that are known for their ability to produce a wide variety of extracellular enzymes, including lignin-degrading enzymes. Streptomyces sp. are commonly found in soil environments and have been shown to degrade lignin in a variety of plant materials.
- Rhodococcus: These are soil-dwelling bacteria that are capable of degrading a wide range of organic compounds, including lignin. Rhodococcus sp. produce a range of lignin-degrading enzymes, including laccases and peroxidases, which enable them to break down lignin.
- Pseudomonas: These are ubiquitous bacteria that are commonly found in soil, water, and other environments. Some strains of Pseudomonas putida have been shown to degrade lignin using a combination of enzymatic and non-enzymatic mechanisms.
- Bacillus: These are spore-forming bacteria that are commonly found in soil environments. Bacillus sp. are known for their ability to produce a variety of lignin-degrading enzymes, including peroxidases and laccases.
- Enterobacter: These are gram-negative bacteria that are commonly found in soil, water, and other environments. Some strains of Enterobacter cloacae have been shown to degrade lignin using a combination of enzymatic and non-enzymatic mechanisms.
Overall, lignin-degrading bacteria are an important component of the microbial community in many environments and play a crucial role in the breakdown of lignin in natural systems. Understanding the diversity and mechanisms of lignin degradation in bacteria can have important implications for biotechnological applications such as bioremediation and lignocellulosic biomass utilization.
2. Lignin-degrading Actinomycetes
Actinomycetes are a group of gram-positive bacteria that are known for their ability to produce a wide range of extracellular enzymes, including lignin-degrading enzymes. Here are some examples of lignin-degrading actinomycetes:
- Streptomyces: Streptomyces sp. are one of the most extensively studied lignin-degrading actinomycetes. They are aerobic bacteria that are commonly found in soil environments and are known for their ability to produce a wide range of lignin-degrading enzymes, including peroxidases and laccases.
- Nocardia: Nocardia sp. are also soil-dwelling bacteria that are capable of degrading lignin. They produce a range of lignin-degrading enzymes, including laccases and peroxidases, which enable them to break down lignin.
- Micromonospora: Micromonospora sp. are another group of lignin-degrading actinomycetes that are commonly found in soil environments. They produce a variety of extracellular enzymes, including lignin peroxidases and manganese peroxidases, which enable them to break down lignin.
- Actinoplanes: Actinoplanes sp. are soil-dwelling bacteria that are known for their ability to produce lignin-degrading enzymes. They produce a range of extracellular enzymes, including laccases and peroxidases, which enable them to break down lignin.
3. Lignin-degrading Fungi
Lignin-degrading fungi are a diverse group of fungi that are capable of breaking down lignin using a variety of enzymatic and non-enzymatic mechanisms. Here are some examples of lignin-degrading fungi:
- White-rot fungi: White-rot fungi are known for their ability to degrade lignin using a range of lignin-degrading enzymes, including lignin peroxidases, manganese peroxidases, and laccases. Some examples of white-rot fungi include Phanerochaete chrysosporium, Trametes versicolor, and Pleurotus ostreatus.
- Brown-rot fungi: Brown-rot fungi are another group of fungi that are capable of breaking down lignin, but they do so using a different set of enzymes than white-rot fungi. Brown-rot fungi primarily degrade the carbohydrates in wood, leaving behind a residue of lignin. Examples of brown-rot fungi include Gloeophyllum trabeum and Postia placenta.
- Soft-rot fungi: Soft-rot fungi are a group of fungi that primarily degrade the cellulose and hemicellulose components of wood, but they can also break down lignin to some extent. Examples of soft-rot fungi include Chaetomium globosum and Serpula lacrymans.
- Endophytic fungi: Endophytic fungi live inside plant tissues without causing any apparent harm to the host plant. Some endophytic fungi are known to produce lignin-degrading enzymes and may play a role in the breakdown of lignin in plant tissues. Examples of endophytic fungi include Phomopsis sp. and Xylaria sp.
The degradation of lignin is an important process in the carbon cycle and is carried out by a variety of microorganisms. Some examples of microorganisms involved in lignin degradation are:
- White-rot fungi: These fungi are known for their ability to degrade lignin using a combination of enzymatic and non-enzymatic mechanisms. Examples of white-rot fungi include Phanerochaete chrysosporium and Pleurotus ostreatus.
- Brown-rot fungi: These fungi are known for their ability to degrade the carbohydrates in wood but have limited ability to degrade lignin. However, recent studies have shown that some brown-rot fungi can also degrade lignin to some extent. Examples of brown-rot fungi include Postia placenta and Gloeophyllum trabeum.
- Bacteria: Several bacterial species are known to degrade lignin, including Streptomyces sp., Rhodococcus sp., and Pseudomonas putida.
- Actinomycetes: These are a group of bacteria-like microorganisms that are known for their ability to degrade lignin. Examples of actinomycetes include Streptomyces sp. and Nocardia sp.
- Ligninolytic basidiomycetes: These are fungi that are able to degrade lignin using a unique set of enzymes called lignin peroxidases. Examples of ligninolytic basidiomycetes include Trametes versicolor and Phlebia radiata.
Lignin is a complex and highly resistant polymer that requires a variety of enzymes to degrade. The enzymes involved in the degradation of lignin can be classified into several groups, including peroxidases, laccases, and oxidases. Here are some examples of enzymes involved in lignin degradation:
- Lignin peroxidases: Lignin peroxidases (LiPs) are heme-containing enzymes that are produced by white-rot fungi and some bacteria. LiPs are able to oxidize and depolymerize lignin through the generation of free radical intermediates. Examples of LiPs include manganese peroxidases (MnPs), versatile peroxidases (VPs), and lignin peroxidases (LiPs).
- Manganese peroxidases: Manganese peroxidases (MnPs) are another group of heme-containing enzymes produced by white-rot fungi. MnPs are able to break down lignin through the oxidation of Mn2+ to Mn3+. Mn3+ is then able to oxidize and depolymerize the lignin polymer. Examples of MnPs include manganese peroxidases (MnPs), versatile peroxidases (VPs), and lignin peroxidases (LiPs).
- Laccases: Laccases are copper-containing enzymes that are produced by a wide range of fungi and bacteria. Laccases are able to oxidize a variety of phenolic and non-phenolic lignin subunits, leading to the depolymerization of lignin. Some examples of laccases include Trametes versicolor laccase and Myceliophthora thermophila laccase.
- DyP-type peroxidases: DyP-type peroxidases are a family of peroxidases that are able to degrade lignin and other aromatic compounds. These enzymes are produced by a wide range of bacteria and fungi and are thought to play a role in lignin degradation in natural systems.
- Oxidases: Oxidases are a group of enzymes that are able to break down lignin by catalyzing the oxidation of lignin subunits. Examples of oxidases involved in lignin degradation include aryl alcohol oxidases, aryl-aldehyde oxidases, and aryl-acid oxidases.
Overall, the degradation of lignin is a complex process that requires the concerted action of a variety of enzymes. The diversity of lignin-degrading enzymes produced by different microorganisms highlights the importance of microbial communities in the breakdown of lignin in natural systems.
Lignin degradation Process
The process of lignin degradation is a complex and multi-step process that involves a variety of enzymes and microorganisms. However, the general steps involved in lignin degradation can be summarized as follows:
- Activation of lignin: The first step in lignin degradation is the activation of lignin, which involves the oxidation of lignin by ligninolytic enzymes such as peroxidases, laccases, and oxidases. This step generates free radical intermediates that are able to break down the lignin polymer.
- Depolymerization of lignin: Once lignin has been activated, the next step is the depolymerization of lignin, which involves the cleavage of the lignin polymer into smaller, more manageable subunits. This step is catalyzed by a variety of enzymes, including peroxidases, laccases, and oxidases.
- Conversion of lignin subunits: The third step in lignin degradation is the conversion of lignin subunits into smaller, more easily metabolizable compounds such as phenols, aromatic acids, and other small organic molecules. This step is carried out by a variety of enzymes, including oxidases, dehydrogenases, and hydrolases.
- Assimilation of lignin degradation products: The final step in lignin degradation is the assimilation of lignin degradation products by microorganisms such as bacteria and fungi. These microorganisms are able to use the carbon and energy stored in lignin degradation products to fuel their own growth and metabolism.
Microbial degradation Mechanisms of lignin
The microbial degradation of lignin is a complex process that involves a variety of mechanisms. Here are some of the key mechanisms involved in the microbial degradation of lignin:
- Oxidative cleavage: One of the primary mechanisms by which microorganisms degrade lignin is through oxidative cleavage. This involves the use of ligninolytic enzymes such as peroxidases and laccases to generate free radical intermediates that are able to cleave the lignin polymer into smaller subunits.
- Reductive cleavage: Another mechanism by which microorganisms degrade lignin is through reductive cleavage. This involves the use of enzymes such as reductases to reduce the quinone and ketone groups present in lignin, leading to the cleavage of the lignin polymer into smaller subunits.
- Hydrolysis: Hydrolysis is another mechanism by which microorganisms can degrade lignin. This involves the use of enzymes such as esterases and hydrolases to break the ester and ether bonds present in lignin, leading to the cleavage of the lignin polymer into smaller subunits.
- Metabolic assimilation: Once lignin has been cleaved into smaller subunits, microorganisms are able to assimilate these subunits into their metabolism. This involves the use of enzymes such as dehydrogenases and oxidases to convert lignin subunits into smaller, more easily metabolizable compounds such as phenols, aromatic acids, and other small organic molecules.
- Synergistic interactions: The degradation of lignin is often carried out by a community of microorganisms that work together in a synergistic manner. This can involve the use of different ligninolytic enzymes by different microorganisms, as well as the exchange of metabolic byproducts that allow microorganisms to more effectively degrade lignin.
- Non-enzymatic reactions: In addition to enzymatic reactions, non-enzymatic reactions can also contribute to lignin degradation. These reactions can be induced by environmental factors such as temperature, pH, and presence of metal ions. Non-enzymatic reactions include radical reactions and Fenton reactions, which involve the generation of reactive oxygen species that can degrade lignin.
- Co-metabolism: Co-metabolism is a mechanism by which microorganisms can degrade lignin in the presence of a more readily metabolizable substrate. For example, microorganisms may use lignin as a source of carbon and energy only when other carbon sources are depleted.
- Transport mechanisms: Once lignin has been degraded into smaller subunits, microorganisms must transport these subunits across their cell membrane to be assimilated into their metabolism. Transport mechanisms such as transporters and permeases are involved in this process.
- Biofilm formation: Microorganisms involved in lignin degradation can form biofilms on lignocellulosic surfaces. Biofilms are communities of microorganisms that adhere to a surface and produce extracellular matrix materials that protect the microorganisms from environmental stresses. Biofilms can enhance the degradation of lignin by creating a localized environment that is conducive to lignin degradation.
- Microbial consortia: The degradation of lignin is often carried out by a consortium of microorganisms that work together to degrade lignin. These consortia can include bacteria, fungi, and actinomycetes, each of which may contribute different enzymes and metabolic pathways to the degradation of lignin.
Examples of microorganisms that can degrade lignin
There are various microorganisms capable of degrading lignin, including:
- White-rot fungi: These fungi are the most effective lignin degraders, as they produce ligninolytic enzymes such as lignin peroxidases, manganese peroxidases, and laccases. Examples of white-rot fungi include Phanerochaete chrysosporium, Trametes versicolor, and Pleurotus ostreatus.
- Bacteria: Certain bacteria are also capable of degrading lignin, including Rhodococcus jostii, Sphingomonas paucimobilis, and Comamonas testosteroni.
- Actinomycetes: Some species of actinomycetes, such as Streptomyces viridosporus and Streptomyces griseus, are known to produce enzymes that can degrade lignin.
- Yeasts: Some yeasts, such as Candida tropicalis and Pichia stipitis, can also break down lignin.
Challenges associated with microbial degradation of lignin
Microbial degradation of lignin presents several challenges, including:
- Complexity of lignin structure: Lignin is a complex polymer made up of different types of monomers and linkages, making it difficult to break down. Microorganisms that can degrade lignin produce a range of enzymes that act on different parts of the lignin structure, but the process is still not fully understood.
- Limited knowledge of lignin-degrading microorganisms: Despite significant progress in identifying lignin-degrading microorganisms, many species and their enzymes remain unknown, and their activity is difficult to measure. This limits our ability to harness their potential for biotechnology applications.
- Competition with other microorganisms: In natural environments, lignin-degrading microorganisms must compete with other microbes for access to lignin, nutrients, and oxygen. This can limit their activity and efficiency in breaking down lignin.
- Toxicity of lignin breakdown products: Lignin degradation can produce toxic intermediates such as reactive oxygen species and aromatic compounds. These can be harmful to microorganisms and other organisms in the environment and can limit the efficiency of the lignin degradation process.
- Limited commercial applications: Despite the potential for lignin degradation to produce valuable products such as biofuels and chemicals, the commercial application of lignin-degrading microorganisms is still limited due to the challenges mentioned above and the high cost of lignin extraction and processing.
What is lignin and why is it important?
Lignin is a complex polymer that provides structural support to plant cell walls. It is the second most abundant organic material on earth after cellulose. Lignin is important because it contributes to the strength and rigidity of plants and is also a potential source of renewable energy and chemicals.
How do microorganisms degrade lignin?
Microorganisms degrade lignin by using ligninolytic enzymes such as peroxidases and laccases to cleave the lignin polymer into smaller subunits. They can also use other enzymes such as esterases and hydrolases to break the ester and ether bonds in lignin.
What factors affect microbial degradation of lignin?
Factors that can affect microbial degradation of lignin include pH, temperature, moisture content, nutrient availability, oxygen availability, and the presence of inhibitors such as heavy metals or pesticides.
What are some examples of microorganisms that can degrade lignin?
Microorganisms that are known to degrade lignin include bacteria, fungi, and actinomycetes. Examples of lignin-degrading microorganisms include Streptomyces viridosporus, Phanerochaete chrysosporium, and Bacillus subtilis.
How can lignin be valorized using microbial degradation?
Microbial degradation of lignin can be used to produce high-value products such as biofuels, bioplastics, and chemicals. By breaking down lignin into smaller subunits, microorganisms can generate feedstocks that can be used for the production of these products.
What are ligninolytic enzymes and how do they work?
Ligninolytic enzymes are a class of enzymes that are capable of breaking down lignin. They work by generating free radical intermediates that can cleave the lignin polymer into smaller subunits.
What are the challenges associated with microbial degradation of lignin?
One of the main challenges associated with microbial degradation of lignin is the complexity of the lignin polymer. Lignin is a highly heterogeneous material that is resistant to degradation. Another challenge is the variability in lignin composition, which can affect the ability of microorganisms to degrade lignin.
How can biotechnology be used to improve microbial degradation of lignin?
Biotechnology can be used to engineer microorganisms that are more efficient at degrading lignin. This can involve the optimization of ligninolytic enzyme production, the enhancement of lignin transport across cell membranes, and the development of synthetic microbial communities that are specialized for lignin degradation.
What are some potential applications of lignin-derived products?
Lignin-derived products can be used in a variety of applications, including as fuel additives, polymer feedstocks, and surfactants. They can also be used in the production of specialty chemicals and materials such as adhesives and carbon fiber.
How does microbial degradation of lignin contribute to the global carbon cycle?
Microbial degradation of lignin is an important process in the global carbon cycle. By breaking down lignin into smaller subunits, microorganisms release carbon back into the ecosystem where it can be used by other organisms. This contributes to the cycling of carbon through terrestrial ecosystems and the atmosphere.
- Crawford, D. L., & Crawford, R. L. (1980). Microbial degradation of lignin. Enzyme and Microbial Technology, 2(1), 11–22. doi:10.1016/0141-0229(80)90003-4
- Janusz, G., Pawlik, A., Sulej, J., Świderska-Burek, U., Jarosz-Wilkołazka, A., & Paszczyński, A. (2017). Lignin degradation: microorganisms, enzymes involved, genomes analysis and evolution. FEMS Microbiology Reviews, 41(6), 941–962. doi:10.1093/femsre/fux049
- Reid, I. D. (1995). Biodegradation of lignin. Canadian Journal of Botany, 73(S1), 1011–1018. doi:10.1139/b95-351
- Geib, S. M., Filley, T. R., Hatcher, P. G., Hoover, K., Carlson, J. E., Jimenez-Gasco, M. d. M., … Tien, M. (2008). Lignin degradation in wood-feeding insects. Proceedings of the National Academy of Sciences, 105(35), 12932–12937. doi:10.1073/pnas.0805257105
- Bugg, T. D. H., Ahmad, M., Hardiman, E. M., & Rahmanpour, R. (2011). Pathways for degradation of lignin in bacteria and fungi. Natural Product Reports, 28(12), 1883. doi:10.1039/c1np00042j
- Atiwesh G, Parrish CC, Banoub J, Le TT. Lignin degradation by microorganisms: A review. Biotechnol Prog. 2022 Mar;38(2):e3226. doi: 10.1002/btpr.3226. Epub 2021 Dec 9. PMID: 34854261.
- Atiwesh, Ghada & Parrish, C. & Banoub, Joseph & Le, Tuyet-Anh. (2022). Lignin Degradation by Microorganisms: A Review. Biotechnology Progress. 38. 12. 10.1002/btpr.3226.
- de Souza, W. R. (2013). Microbial Degradation of Lignocellulosic Biomass. InTech. doi: 10.5772/54325