1. Introduction
Many industries currently produce many tons of agro-industrial wastes. However, direct utilization of lignocellulosic biomass as a feedstock for bioproducts is challenging due to their complex structure (as represented in
Figure 1). A variety of useful components, including sugars, protein, lipids, cellulose, and lignin, are present in natural fibres. The major issue that limits their utilization is, however, the tight bonding within their components
[1]. Cross-linking of polysaccharides and lignin occurs through ester and ether bonds, while microfibrils produced by cellulose, hemicellulose, and lignin aid in the stability of plant cell wall structure
[2][3]. These strong cross-linking connections exist between the components of the plant cell wall that act as a barrier to its disintegration.
Figure 1. Overview of the complex structure of natural fibers and pretreatments.
Pretreatment helps to fractionate biomass prior to further processes, making it simpler to handle in the process
[4][5][6]. It enables biomass hydrolysis and makes building blocks for biobased products, fuels, and chemicals. It is often the initial stage of the biorefining process and enables the following steps such as enzymatic hydrolysis and fermentation to be carried out more quickly, effectively, and economically
[7]. The pretreatment method used is entirely dependent on the targeted application. Numerous pretreatment methods are mainly developed to effectively separate these interconnected components in order to get the most advantages from the lignocellulosic biomass’s constituents.
Pretreatment of natural fibres is not as straightforward as it may seem. In fact, it is the second most expensive procedure after the installation of a power generator. Hydrogen bond disruption, cross-linked matrix disruption, as well as increased porosity and surface area, are the three objectives that a good pretreatment technique accomplishes in crystalline cellulose. Additionally, the result of pretreatment varies attributed to the different ratios of cell wall components
[8]. More criteria to take into consideration for efficient and economically feasible pretreatment process include less chemical usage, prevention of hemicellulose and cellulose from denaturation, minimum energy demand, low price, and the capacity to reduce size.
Biomass recalcitrance is a term used for the ability of natural fibres to resist chemical and biological degradation. While there are many components involved in the recalcitrance of lignocellulosic biomass, the crystalline structure of cellulose, the degree of lignification, accessible surface area (porosity), the structural heterogeneity, and complexity of cell-wall are primary causes
[9][10]. As a consequence of breaking the resistant structure of lignocellulose, it causes lignin sheath, hemicellulose, and crystallinity to all be degraded, as well as casuing a decrease in cellulose’s degree of polymerization
[11].
Depending on the types of natural fibres employed, the preference for the pretreatment method varies according to the composition of cellulose, hemicellulose, and lignin.
Figure 2 depicts the general differences between the many common approaches which come under the four categories of physical, chemical, biological, and combination pretreatment
[4]. While some of these methods have successfully transitioned from a research platform to an industrial stage, there are many hurdles, and one of the greatest is the requirement for highly toxic waste generation and high-energy inputs. From here, a serious issue that must be addressed is the lack of green and cost-effective solutions. Nevertheless, it has only lately garnered significant attention as a potential solution to the problem by focusing on the employment of non-chemical pretreatment. This could be reflected by the increment in article publications that reviewed lignocellulosic fibre pretreatment via individual greener approach as highlighted in
Table 1 indicating that this topic is increasingly well-known owing to environmental concerns. The development of technology that maximises the use of raw resources, reduces waste, and avoids the use of poisonous and hazardous compounds is critical to accomplishing this objective. However, a review of all greener pretreatment approaches for lignocellulosic biomass is missing in the current literature.
Figure 2. Different pretreatments, which fall into four main categories: physical, chemical, biological, and combination have been used to improve lignocellulosic fractionation for natural fibres.
Table 1. Recent review articles related to greener pretreatment approaches for lignocellulosic biomass.
| No. |
Title |
Highlights of Review |
Ref. |
| 1. |
Enzymatic pretreatment of lignocellulosic biomass for enhanced biomethane production-A review |
-
Reviews the anaerobic digestion process, challenges in degrading lignocellulosic materials, the current status of research to improve the biogas rate and yield from the anaerobic digestion of lignocellulosic biomass via enzymatic pretreatment, and the future trend in research for the reduction of enzymatic pretreatment cost
|
[12] |
| 2. |
A review on the environment-friendly emerging techniques for pretreatment of lignocellulosic biomass: Mechanistic insight and advancement |
|
[13] |
| 3. |
Recent Insights into Lignocellulosic Biomass Pyrolysis: A Critical Review on Pretreatment, Characterization, and Products Upgrading |
|
[14] |
| 4. |
Recent advances in the pretreatment of lignocellulosic biomass for biofuels and value-added products |
|
[15] |
| 5. |
Emerging technologies for the pretreatment of lignocellulosic biomass |
|
[16] |
Hence, the green pretreatment approaches for lignocellulosic biomass such as physical, biological, and combination methods, as well as their impact on the separation of the complex components of different lignocellulosic sources, are reviewed in more detail in the next sections.
3. Biological Pretreatment
Retting is a biological process in which enzymatic activity removes non-cellulosic components connected to the fibre bundle, resulting in detached cellulosic fibres. The dew retting uses anaerobic bacteria fermentation and fungal colonization to produce enzymes that hydrolyse fibre-binding components on fibre bundles.
Clostridium sp. is an anaerobic bacterium commonly found in lakes, rivers, and ponds. Plant stems were cut and equally scattered in the fields during the dew retting process, where bacteria, sunlight, atmospheric air and dew caused the disintegration of stem cellular tissues and sticky compounds that encircled the fibres
[23]. For the dew retting procedure to enhance fungal colonization, locations with a warm day and heavy might dew are recommended.
Bleuze et al.
[24] investigated the flax fibre’s modifications during the dew retting process. Microbial colonization can be affected the chemical compositions of cell walls. After seven days, fungal hyphae and parenchyma were found on the epidermis and around fibre bundles, respectively. After the retting process (42 days), signs of parenchyma deterioration and fibre bundle decohesion revealed microbial infestation at the stem’s inner core.
Fila et al.
[25] found 23 different varieties of dew-retting agent fungi in Southern Europe. All
Aspergillus and
Penicillium strains yield high-quality retted flax fibres, according to the researchers. Besides that, under field conditions, Repeckien and Jankauskiene
[26] investigated the effects of fungal complexes on flax dew-retting acceleration.
Cladosporium species variations with high colonization rates (25–29%) have been identified as a good fungus for fibre separation. Most fungi survived on flax fed with fungal complex N-3, which contained six different fungal strains.
On a commercial scale, Jankauskiene et al.
[27] optimised the dew retting method. Two fungal combinations were created and put to straw after the swath was pulled and returned. Furthermore, after spraying
Cladosporium herbarum suspension during fibre harvesting, extremely high fibre separation was found.
Bacterial and Fungi Interaction
Fungi colonization is thought to be the most important enzymatic active mechanism for dew retting. Recent research has focused on the interplay of the bacterial and fungal communities during dew retting. The association between the chemical contents of hemp fibres and microbial population fluctuation during the retting process was investigated by Liu et al.
[28]. In the first seven days, fungal colonization was discovered with very little bacteria. After 20 days, there was a gradually risen in bacterial attachments on the fibre surface, with fewer fungal hyphae. The area with the highest bacterial concentration was found to severely deteriorate. The phylogenetic tree for the bacterial and fungal population in dew-retting hemp fibres is shown in
Figure 3. While
Table 2 shows ultrastructural changes in hemp stems and fibres as a result of microbial activity during the retting process.
Figure 3. The phylogenetic tree of the (
a) bacterial and (
b) fungus communities found in hemp fibre samples. The color of the branches indicates the type of proteobacteria present, while the color of the tag indicates the number of bacteria/fungi present on different days
[28].
Table 2. Highlights of ultrastructural changes on hemp stems and fibres associated with microbial activity during the retting process
[29].
| Retting Period |
0 Days |
7 Days |
14–20 Days |
After 50 Days |
| Changes in the hemp stem’s and fibre’s ultrastructure |
(i) Stem with a well-preserved layered structure (ii) Un-collapsed, unbroken cells with their original cell geometry (iii) Living cells with cytoplasm (iv) Cuticle and trichomes are unharmed on the clear surface. (v) Chloroplasts in abundance in the upper epidermis |
(i) The structure as a whole is in good condition. (ii) Fungal growth on the outside of the stems and inside the stems (iii) With damaged epidermis and parenchyma, cellular architecture is less stable. |
(i) Cuticle has seriously deteriorated. (ii) Changes in cellular anatomy, as well as significant loss of live cells (iii) Fibre bundles were isolated from each other and the epidermis. (iv) Thick-walled cells populate seldom; parenchyma degrades completely, although chlorenchyma suffers less harm. (v) Bast fibres with sporadic moderate attacks (vi) Fungi colonisation and decay morphology were both affected by fibre morphology. |
(i) The structure of hemp was severely harmed and dissolved. (ii) The epidermis and cambium were heavily invaded by dominating bacteria. (iii) In the bast regions, the parenchyma cells have been destroyed, and the structural integrity has been lost. (iv) All cell types, including fibre cells, have hyphae inside their lumina. (v) BFIs are more intense inside the stem. (vi) Anatomy and ultrastructure have been severely harmed. (vii) Bast fibres with a thick wall and degradation properties (viii) Effects on the ultrastructure of the fibre wall.
-
CML loosening/degradation, resulting in delamination and defibration
-
The S3 layer is loosening and decaying
-
Delamination within the S2 trans wall and intra wall cracks in the S2 layer have a noticeable effect
-
S2 materials have been removed directly (e.g., S2 thinning, broken S2, and disintegration into nanosized cellulose fibrillar structures)
|
| The dynamics and activity of microbes |
Fungi (i) Rarely seen Bacteria (ii) Not observed Fungi |
Fungi (i) Mycelia with sparse growth (ii) Less variety (iii) Outside of the cortical layers, colonisation occurs largely in live cells. (iv) Trichomes near to the surface trichomes have dense colonisation. (v) Dependence on readily available food (vi) Damage to cell walls is reduced. Bacteria (i) Less abundant |
Fungi (i) Extensive and plentiful (ii) Mycelia densely covering the cuticle (iii) diverse population (iv) a large number of spores (v) Interactions and activities that are intense Bacteria (i) Abundant (ii) Diverse population iii) Over the cuticle, colonies (iv) Associated with hyphae and fungal spores (v) After 20 days, there are more noticeable activity (vi) Cuticle has severely deteriorated |
Fungi (i) Less abundant on the outside of the stem (ii) Mycelia on the surface is dead, but there are active hyphae inside the stem (iii) Mycelia, an invading bacteria’s sole source of nourishment, showed bacterial mycophagy (i.e., extracellular and endocellular biotrophic and extracellular necrotrophic activities). Bacteria (i) Highly abundant inside and outside the stems (ii) Highly dominant and diverse role. (iii) Visible as dense overlay representing (a) Biofilms (b) Morphologically different colonies (c) Randomly scattered cells (iv) Showed strong BFIs (v) Using fungal highways, bacterial movement occurs over and inside the hemp stem. (vi) Cutinolytic and cellulolytic activities were improved. |