Different Aspects of Chemical and Biochemical Methods for Chitin Production a Short Review
Broquá J, Zanin B.G, Flach A.M, Mallmann C, Taborda F.G.D, Machado L.E.L, Alves S.M.L, Silva M.M, Dias R.J.S.P, Reis O. V, Pighinelli L*
Department of BIOMATTER, Lutheran University of Brazil (ULBRA), Brazil
*Corresponding author: Luciano Pighinelli, Department of BIOMATTER, Lutheran University of Brazil (ULBRA), Brazil. Post- Graduation Graduate Program in Genetics and Toxicology (PPGGTA), Biomatter Lab, Lutheran University of Brazil (ULBRA), Canoas, Brazil. Tel: +5551995656905; Email: pighinelli@gmail.com
Received Date: 17 April, 2018; Accepted Date: 01 May, 2018; Published
Date: 08 May, 2018
Citation: Broquá J, Zanin BG, Flach AM, Mallmann C,
Taborda FGD, et al. (2018) Different Aspects of Chemical and Biochemical
Methods for Chitin Production a Short Review. J Nanomed Nanosci: JNAN-145. DOI:10.29011/2577-1477.100045
Abstract
The importance of chitin
and its derivatives has grown significantly over the last two decades due to
its renewable and biodegradable, biocompatible source, and also because of the
increase in the knowledge of its functionality in the technological and
biomedical applications.The present short review the biopolymer chitin
and its derivatives as versatile biomaterials for many applications such
pharmaceutical, food, chemistry, agribusiness and medical area, also shows the
main limitations of the research groups and the industry to work with this
biomaterial. This short review tries to compare many aspects of the production
with chemical and biochemical process using different bio catalyser to produce different
characteristics of the chitin and its derivatives. In the final considerations,
the chitin and its derivatives have a great potential to design new
biomaterials for many applications, depends the different characteristics of
the process such, chemicals, concentration, time, temperature, bio catalyser,
that could affect in the structure of the material reflecting directly with
different properties, reducing the costs of the production, less time and
making this biopolymer more attractive for the industry.
Introduction
In addition to being the second most abundant
natural polymer in nature, a renown has a great potential as a biomaterial in
the area of biotechnology, because it is biocompatible, bioreactive and
biodegradable [1]. Such characteristics, diverse applications in areas such as
agriculture, food, environmental, and as two areas with greater focus:
pharmaceutical and health [2,3]. Its structure consists of
N-acetyl-d-glucosamine units with β- (1,4) bonds, having as main characteristic
the insolubility in water and some organic acids [4]. Chitin belongs to the
group of structural polysaccharides, together with cellulose, the second
polymer being more abundant in the biosphere [5-6]. Due to its structural
nature, a product release system was not found in any of the arthropod
exoskeleton, in the structures of Molluscs [7], in the cell wall of fungi [7-8],
protozoa and bacteria, egg shells of nematodes [8], the shrimp fishery residue
being the most widely used source [9]. Throughout the decades of research and
handling of this polymer, many methods of extraction have been developed, being
the chemical method, most found in the literature, being also used in the means
of production of industrial chitin. The USA, Japan, India, Canada, China, South
Korea, Russia and Norway generally use the reject of crustacean fishing for
production. The use of strong acids and bases in the chitin extraction process
generates critical points to the process, such as: high cost of the materials involved,
generation of chemical effluent and final product with low levels of purity [5,10].
Biological processes become more attractive because they have an affordable
cost of production, do not generate high risk effluent (such as the chemical
process) and high quality final product [1,10]. All the processes found in the
literature are an objective: to obtain chitin by separating the proteins and
minerals from the raw material used [2].
Chitin, besides having great biotechnological
value, generates by-products (such as chitosan) that also have added value and
even more relevant properties. In this paper we discuss the already known
processes of obtaining chitin known and registered in the literature of 2010 up
to the present moment: Chemical, enzymatic and biological processes relating
the different methods of obtaining and with the objective to identify the
particularities of each process regarding the industrial viability and
economically balancing them so that the reader concludes the best process for their
research, also the possibility of executing quality improvements in these
processes. We will also discuss the polymorphic structures of α- and β-chitin
and the different methods of obtaining each, since different processes are
required in each of them due to their structures, properties and reactivity.
The main objective of this review is to be able to relate the different
processes of obtaining chitin with the most suitable applications for the
method, based on such relation in aspects such as degree of purity and economic
applicability.
Chitin
Economic
Aspect
The main industrial source used for the
extraction of the biopolymer is the reject of the fishing industry, reusing
mainly the shell of prawns, crabs and lobsters [11-12]. According to Ioelovich
(2014), the production of chitin and its derivatives is estimated at 100
billion tons per year [13]. Gortari & Hours (2013) states that the bark
discarded by the fishing trade can reach 70% of the total weight of the
material [14]. According to Ameh et al. (2014), these carapaces have about
20-30% chitin, varying according to species and season, since chitin is
naturally produced on a scale of 1010 to 1011 tons each
year in a study conducted by Ifuku, et al. (2015) [5,15]. Jaganathan, et al.
(2016) estimates that chitin costs $ 220 per kilogram on the world market [11].
Chemical
Aspect
Chitin is a natural polymer composed of units
of β-D-glucosamine molecules and N-acetyl groups, forming
monomers that will be bound by beta (1 → 4) bonds [2]. Its Degree of
Acetylation (DA) must be above 50% to be characterized as chitin [16]. Thus, it
is called β (1 → 4) -N-Acetyl-D-glucosamine (Figure 1) [5,17]. The
polymer should have acetylation degree greater than 50% to be called chitin Due
to the beta bonds between carbons 1 and 4, chitin becomes an extremely stable
polymer [18]. Its polymeric extension renders it insoluble in water and
practically all organic acids due to the intense hydrogen bonds [19-21]. Chitin
contains, on average, 6.5% of nitrogen and its main derivative, chitosan, can
reach up to 9.5% of hydrogen content. In obtaining the chitosan, the polymer
undergoes a deacetylation process, eliminating about 80% of the acetyl groups
to obtain the amino group [22].
Chitin has statistically ordered allomorphic configurations. These arrangements, or microfibres, can be characterized in: α-chitin, β-chitin and γ-chitin (Figure 2) [8]. Α-Chitin has an antiparallel arrangement of its microfibers that favor hydrogen bonds and give rise to a highly compact structure, resulting in high crystallinity and hardness. It is also known as the most abundant form in nature [18]. Β-chitin has parallel chains, being less crystalline and with less packaging, being more flexible and more reactive. Γ-chitin is a mixed composition of forms α and β [6,10]. Α-chitin can be transformed into β-chitin, but not the reverse [23]. The three allomorphs can be observed by X-ray diffraction and Nuclear Magnetic Resonance (NMR). The degradability of the polymer occurs through the action of the enzyme chitinase, present in nature [24]. Of course, chitin is found in the macrofibrillar form, allowing the production of nanofibers by mechanical or chemical processes, according to Azuma, K. et al. (2014) [25].
Because it is a natural polymer, chitin has
characteristics characteristic of polysaccharides, such as: biocompatibility,
biodegradability and non-toxicity. It still has bioactivity, antibacterial
activities [26] and antimicrobial [25], anticancer activity, anticoagulant [27]
and molecular adsorption property [17-18]. Elieh Ali Komi, Sharma and Dela Cruz
(2017) highlights the immunogenic property of chitin and chitinase against
pathogens in both insects and mammals as well as plants [8]. It is physically
versatile, can be processed in the form of powder, fibers, membranes, sponges,
hydrogels [22], scaffolds [21,28]. According to Anitha et al. (2014), the
structure of sponges is very favorable for health, with emphasis on tissue
engineering, as it has a hygroscopic potential and complexing capacity of other
substances to the structure [29].
Applications
Amidst all its unique properties, chitin and its derivatives are an excellent biomaterial with unlimited application possibilities. From the chemical industry [30-31] and agrochemicals [2] to the textile industry [1,5,26,31-34,] and paper industry [13,26,35,] chitin and derivatives are used as biomaterials potential. Most of the studies found in the literature are directed to the health area, as noted below in Table 1. Kandra, Challa and Kalangi Padma Jyothi (2012) presents a vast study on the performance of chitin and chitinase in immunology [22]. Biocomposites of chitin and mainly chitosan is widely studied and developed [7,13,20]. Chitosan, because of its greater bioactivity and adhesiveness, becomes more interesting as scaffolds for areas of medicine and pharmacy and for tissue engineering [29].
The Major
Limitation of Chitin Production
The application of
chitin and its derivatives depends upon the useful form of the copolymer for
different places to use and the quality of the structural and amorphous part of
the polymers involved.
The major limitation in the use the chitin and chitosan for design applications are related with: [36-37].
·
The
collection of the raw material; different characteristics of the initial
structure such Mw, ash content and the selection of Alpha or Beta chitin;
·
Difficult
to obtain reproducible products with different raw materials;
·
The
cost of production still high;
·
The
absence of validated process and products of biopolymer manufacture;
·
No
standardization of product quality and product assay methods for chitin and
chitosan.
Extraction and Characterization of Chitin
For decades, chitin has been studied in order
to optimize its biomaterial production methods and to better understand its
properties and biomaterial potential. Currently, there are several methods of
obtaining them in the literature, being the biological processes, with the use
of microorganisms and enzymes [1,5,10-11], chemical [6,38] and even a
combination of both to achieve an even more process efficient, as published by
Younes et al. 2012 [30]. Regardless of the process adopted, the goal is the
same: eliminate proteins, minerals, lipids and pigments until only the material
of interest is obtained. Today, the most common method in industrial production
is the chemical method, due to its productivity and practicality, however, all
forms of obtaining chitin are relatively accessible and quite practical [39].
Chemical
Process of Obtaining
This is currently the most widely used method in both industrial and laboratory production and is also the most frequently cited in the literature [40]. As mentioned earlier, the purpose of any extraction process is to eliminate all organic and mineral content of the raw material. For this, there are two primordial steps to obtain chitin: deproteinization and demineralization [38]. The order of these steps in a process can be changed according to the purpose and chemicals used. In addition to these two main steps, the production can receive a step of depigmentation and deodorization, if necessary [41].
Firstly, the raw material receives a preliminary treatment to remove impurities and coarse organic waste. At this stage, baths are carried out with deionized water and sodium hypochlorite is sometimes applied [14]. The use of temperature is eventually employed to speed up the cleaning process. The material is still dry or not to follow the milling step, where it will provide a better reaction with subsequent steps [41]. All types of pretreatment are adopted according to the need and condition of the waste used, as well as the species of selected raw material [18].
Deproteinization is the elimination of the protein content present in the raw material. For this, alkaline solutions are used, for example NaOH and KOH [6]. The most used and adopted for industrial production is NaOH solution [42]. A large variation in production aspects is found in the literature. Gortari and Hours, 2013 shows that temperatures in the average of 95ºC are used in the deproteinization for commercial chitin, but also indicates that in this range can cause the depolymerization of the material and change some characteristics such as viscosity [14].
Demineralization involves the removal of the inorganic filler (calcium carbonate and calcium phosphate) from the raw material [43]. For this, inorganic acids, such as HCl, HNO₃ and H₂SO₄, and also strong organic acids are used, for example: HCOOH and CH₃COOH [7]. The most commonly used acid in the production of commercial chitin is HCl, due to its high efficiency in the removal of the minerals present. The staining is a strong indicator of the presence of impurities in the material. For this, the depigmentation step is carried out using acetone, sodium hypochlorite, hydrogen peroxide or potassium permanganate [7,27]. Table 2 shows a comparison between some chemical processes found in the literature.
Biotechnological
Process of Obtaining
The biotechnological process is the
combination of the chemical process with the use of biological methods, with
application of microorganisms to the system [43]. This technique shows to be
favorable in comparison with the traditional chemical process, where large
amounts of highly reactive chemical inputs are used, which can affect the final
quality of the material, besides the serious effluent generated in the process.
Methods involving biological pathways have also been shown to be more assertive
when achieving higher purity states of chitin, with considerably lower
molecular weight loss than the conventional chemical process [33-34]. Chemical
processes still prove to be more efficient industrially, however the use of
biotechnological methods presents a new sustainable vision and new parameter of
quality, offering a more suitable biomaterial for health areas [10].
Demineralization can be performed using microorganisms. The material is deposited along with a microbial culture and a sugar source, which will provide the necessary nutrients. As a result of this fermentation process, there are the production of organic acids that react with the minerals and turn them into salts and precipitate. At the end of the process, the above can be removed with a simple washing process [1,27]. According to Aranday-García et al. (2017), lactic acid from the activity of lacto bacteria, provides better results and greater efficiency in demineralization [44]. The inoculation takes place according to each treated species in a culture medium with the necessary nutrients. This medium is basically composed of sugars and fats; amino acids; sources of calcium, iron and magnesium; among other specific compounds. Agar, commercial culture media, are the most used for presenting balanced composition, such as MRS (Man Rogosa Sharpe) [45].
For the elimination of proteins, fermentative and enzymatic processes can be applied. During fermentation, digestive and microbial enzymes are produced and consume organic material [44]. Hydrolytic enzymes (proteases) are very efficient in deproteinization and may result in the production of hydrolyzed proteins as a by-product of high added value [32,46]. Proteases from lacto bacteria are the most commonly used [39]. Carotenoids are pigments found in lipids present in crustaceans and can be isolated from fermentation and enzymatic activity [47]. Astaxanthin belongs to these carotenoids present and has great commercial value in the food area [32].
The main disadvantage of processes involving microorganisms are the time released for fermentation and high cost of some enzymes [40,48]. Often the biological process is insufficient, requiring the application of acids and alkaline solutions to confirm the reaction, but presenting a better deproteinization yield, as described in Table 3, consequently obtaining a material of significantly higher quality due to low molecular weight loss [45].
Final Considerations
The mastery of the techniques and study of the kinetics of each process is fundamental for the optimization of the same. The analysis of the different chitin obtaining studies by chemical means found in the literature suggests a higher efficiency in the use of consecutive and short baths instead of a long one for deproteinization as well as for the demineralization, thus dispensing with the use of solutions with large concentrations of reagents, which in turn avoids the loss of the quality of the chitin obtained in the process. Obtaining by biotechnological methods presents better results in most literary studies, however, it lacks further development, since the average time in the process is often greater than by the chemical method, leaving the employment of bacteria and enzymes less attractive for the chitin production at industrial levels. Of these bacteria, a family that proves very promising is Lactobacillus, because in the process this produces lactic acid, simultaneously promoting the demineralization of the material.
Chitin and its derivatives,
as previously mentioned, have several applications in the most varied areas,
but the current trend of research seems to be more focused on the biomedical
area, with the development of treatments and methods that promote the
regeneration of wounds and / or nervous tissues. For the application of chitin
in these areas it becomes more necessary to use a material with a higher degree
of purity, so it is advisable to use biotechnological methods in these cases.
The fact that the degree of purity is lower in chemical processes does not
deprive the application of the chitin obtained by them in other areas that do
not demand such purity, these areas may be more attractive for investments,
taking advantage of the faster production and greater amount provided by such
methods.
Molecular representation of (A) cellulose, (B) chitin and (C) chitosan.
Figure 1:
Difference between cellulose, chitin and chitosan.
Figure 2: Chitin
allomorphs.
Area |
Applications |
References |
Agriculture |
Soil modifier; fungicide [6]; defensive mechanism in plants; seed
coating; time release of fertilizers [17]; agro-bioscience [49]. |
[1,5,13,34] |
Biocatalysis |
Scaffolds as a support for metals in order to produce catalysts [29] |
[35] |
Bioengineering |
Reduced susceptibility to echinocandin with elevated chitin levels in C. albicans cells [50] |
[29, 50] |
Biomedical |
Enzyme immobilization and purification chelator [8-9]; emulsifier;
flocculen [6]; blood cholesterol control [23,26]; Lectin affinity
chromatography; biosensor [48] immobilization of antibody in the presence of
alginate [8]; haemostatic agents [29]. |
[23,45,49] |
Biotechnological products |
Packaging [32]; manufacture of film and sponge sheet materials [26]; hydrogels [28]. |
[14,18] |
Cosmetic |
Lotions; hair additives; body creams [6]. |
[5,31-34,51] |
Energy production |
Clostridium
paraputrificum M-21 to produce hydrogen gas as potential source of
alternative energy [22] |
[22] |
Food industries |
Stabilizing agent [9]; dietary supplements [1]; antioxidant [6];
emulsifying agent [35]; food preservation such as for edible films [27];
weight loss agent; food and feed additives [26]. |
[31-34,49,51] |
Medical |
Fibers; membranes; artificial organs [6] and skin; surgical sutures [29];
bone and cartilage regeneration [27]; wound healing and dressings; cancer
diagnosis [8,24]; aid in cataract surgery; periodontal disease treatment [26];
collagen synthesis [28]; contact lenses [29]; tumor therapy; stem cell
tecnology [24]. |
[21,27-28,52] |
Pharmaceutical |
Manganese supplement complex [3]; drug release [6,18]; gene delivery [24]. |
[2,9,27,45,53-54] |
Pollutants removal |
Copper removal capability to obtain more stable diesel oil. |
[22] |
Water treatment |
Dye removal [17]; absorbent for heavy and radioactive metals [26]. |
[13,26,33] |
Table 1:
Applications of chitin and its derivatives as a biomaterial.
Source |
Deproteinization |
Condition |
DP % |
Demineralization |
Condition |
DM % |
Ref |
|
Chicken feet |
1:10 (w/v) at 1 N NaOH solution |
24h, 90°C |
NI |
1:18 (w/v) at 1.5N HCl solution |
36h, RT |
NI |
[26] |
|
Crab Shells |
- |
- |
- |
1:10 (w/v) at 0,55M HCl solution |
15 min, 4 baths, RT |
99 |
[06] |
|
Crustacean waste¹ |
1:60 (w/v)² at 1.25 M NaOH solution |
2h, 90°C,
stirring |
~78 |
1:10 (w/v) at 1M HCl solution |
60 min, RT, 800 rpm |
98 |
[34] |
|
1:60 (w/v)² at 1.25 M NaOH solution
(seawater with 35% salinity as base) |
2h, 90°C,
stirring |
~76 |
1:10 (w/v) at 1M HCl solution (seawater
with 35% salinity as base) |
60 min, RT, 800 rpm |
89 |
|||
Cuttlefish |
- |
- |
- |
1:10 (w/v) at 0,55M HCl solution |
15 min, 4 baths, RT |
98 |
[06] |
|
Fish Scales |
n/c (w/v) at 2N NaOH solution |
36h, RT |
NI |
n/c (w/v) at 1% HCl solution |
36h, RT |
NI |
[38] |
|
Housefly pupa Shells |
1:11 (g/mL) at 1.25 N NaOH solution |
3h, 95°C |
NI |
1:11 (g/mL) at 2N HCl solution |
3h, RT |
NI |
[55] |
|
Mushroom |
n/c (w/v) at 2M NaOH solution |
24h, 85°C |
NI |
1:12.5 (w/v) at 2M HCl solution |
15h, 60°C |
NI |
[42] |
|
Shrimp |
- |
- |
- |
1:8 (w/v) at 0.2M C₃H₆O₃ solution |
20 min, RT, Stirred |
65 |
[05] |
|
1:8 (w/v) at 1M C₃H₆O₃ solution |
20 min, RT, Stirred |
97 |
||||||
Shrimp |
- |
- |
- |
1:10 (w/v) at 0,55M HCl solution |
15 min, 4 baths, RT |
99 |
[06] |
|
Shrimp |
1:25 (g/mL) at 0,5M NaOH solution |
2h, RT |
71³ |
1:25 (g/mL) at 0,5M HCl solution |
2h, RT |
80³ |
[17] |
|
Shrimp |
- |
- |
- |
1:10 (w/v) at 2M HCl solution |
90 min, 50°C,
30 rpm, 3 baths |
100 |
[39] |
|
1:10 (w/v) at 0.5M HCl solution |
90 min, 4°C,
30 rpm, 3 baths |
|||||||
Shrimp |
- |
- |
- |
1:10 (w/v) at 1,5M HCl Solution |
6h, 50°C
150 rpm |
98 |
[30] |
|
Shrimp |
- |
- |
- |
n/c (w/v) at 1.1 M C₃H₆O₃ solution |
20 min, 25°C |
99 |
[40] |
|
Shrimp |
- |
- |
- |
1:10 (w/v) at 1.1M HCl solution |
6h, 20°C,
300 rpm |
99 |
[51] |
|
1:10 (w/v) at 1.5M CH₂O₂ solution |
6h, 20°C,
300 rpm |
99 |
||||||
1:10 (w/v) at 2.35M CH₃COOH solution |
6h, 20°C,
300 rpm |
99 |
||||||
1:10 (w/v) at 1M C₆H₈O₇ solution |
6h, 20°C,
300 rpm |
99 |
||||||
1:10 (w/v) at 1.5M H₃PO₄ solution |
6h, 20°C,
300 rpm |
99 |
||||||
NI: Not informed (¹):
Crustacean waste was composed of a variety of crustacean's shells |
Table 2:
Chemical process parameters.
Microrganism |
Process |
Substrate |
Efficiency |
Ref |
||||||||
|
Temp (°C) |
Stirring (RPM) |
Time (h) |
|
DM % |
DP % |
Chitin % |
|
||||
Lactobacillus sp. B2 |
Fermentation (DM and DP occur
simultaneously at an orbital shaker) |
30 |
200 |
120 |
|
88 |
56 |
34 |
[01] |
|||
B. mojavensis A21 |
Enzymatic DP under alkaline solution (4%
NaOH) |
50 |
- |
3 |
|
- |
89¹, 94²
and 70³ |
37¹, 27²
and 6³ |
[06] |
|||
P. aeruginosa |
Fermentation aiming production of organic
acid in orbital shaker |
30 |
180 |
168 |
D- Glucose 10% (w/v) |
~91 |
- |
From 11,56 to 44,25⁴ |
[11] |
|||
S. marcescens |
Fermentation aiming DP in orbital shaker |
30 |
180 |
168 |
- |
NI |
||||||
L. plantarum PTCC 1058 |
Fermentation in incubator shaker (DM and DP
occur simultaneously) |
30 |
180 |
144 |
Date syrup |
54 |
45 |
NI |
[17] |
|||
B. cereus SV1 |
Fermentation (DM and DP occur simultaneously) |
37 |
200 |
120 |
Glucose 5% (w/v) |
77,3 |
88,6 |
- |
[12] |
|||
B. subtilis A26 |
79,9 |
91,25 |
- |
|||||||||
B. mojavensis A21 |
78,7 |
88 |
- |
|||||||||
B. pumilus A1 |
75,3 |
91,2 |
- |
|||||||||
B. lichenformis RP1 |
55,55 |
90,8 |
- |
|||||||||
B. amyloliquefaciens An6 |
66,05 |
90,8 |
- |
|||||||||
B. mojavensis A21 |
Enzymatic DP |
50 |
NI |
3 |
NI |
- |
77 |
NI |
[39] |
|||
B. subtilis A26 |
40 |
- |
75 |
|||||||||
V. metschnikovii J1 |
40 |
- |
75 |
|||||||||
B. lichenformis MP1 |
50 |
- |
75 |
|||||||||
B. lichenformis NH1 |
50 |
- |
65 |
|||||||||
A. clavatus ES1 |
40 |
- |
59 |
|||||||||
Scorpaena scrofa (scorpion fish) enzyme |
45 |
- |
80 |
|||||||||
B. mojavensis A21 |
Enzymatic DP |
50 |
NI |
NI |
|
- |
76 |
NI |
[30] |
|||
V. metschnikovii J1 |
40 |
- |
||||||||||
B. lichenformis MP1 |
50 |
- |
||||||||||
B. subtilis A26 |
40 |
- |
||||||||||
B. lichenformis NH1 |
50 |
- |
65 |
|||||||||
A. clavatus ES1 |
40 |
|
- |
59 |
||||||||
Alcalase (Commercial enzyme) |
50 |
No substract |
- |
54 |
||||||||
Bromelain (Commercial enzyme) |
50 |
- |
67 |
|||||||||
S. griseus |
Enzymatic DP |
37 |
0 |
5 |
KH₂PO₄ (0,5M/L) |
- |
91 |
NI |
[40] |
|||
S. thermophilus |
Fermentation in incubator (DM and DP occur
simultaneously) |
42 |
0 |
72 |
Glucose 15% (w/v) |
92 |
94 |
NI |
[33] |
|||
B. subtilis⁵ |
Fermentation in rotary shaker (DM and DP
occur simultaneously) |
60 |
150 |
24 |
Glucose 1% (w/v) |
94⁶ |
84 |
NI |
[34] |
|||
B. licheniformis and |
Fermentation in incubator shaker (DM and DP
occur simultaneously) |
30 |
180 |
60 |
No substract |
93.5 |
87 |
NI |
[45] |
|||
G. oxydans⁷ |
36 |
Glucose 5% (w/v) |
||||||||||
L. brevis |
Fermentation (DM and DP occur
simultaneously) |
30 |
0 |
192 |
Glucose 10% (w/v) with Rhizopus oligosporus spores⁸ |
67,3 |
96 |
NI |
[44] |
|||
Portunus segnis (blue crab) viscera enzyme, Purafect (R 2000E) and Neutrase (P1236) |
Enzymatic DP |
50 |
0 |
3 |
No substract |
- |
91 |
22,2 |
[31] |
|||
P saliphilus |
Enzymatic DP |
50 |
0 |
168 |
|
- |
85.6 |
NI |
[46] |
|||
A. niger |
Enzymatic DP under acid solution |
20 |
300 |
6 |
No substract |
- |
95 |
NI |
[51] |
|||
NI: Not informed |
⁴: Yield varies according to crustacean source |
⁸: Substrates
added after 120h of fermentation |
||||||||||
¹: Yield from shrimp shells |
⁵: Medium using seawater |
|
||||||||||
²: Yield from crab shells |
⁶: DP attained after further chemical treatment |
|
||||||||||
³: Yield from cuttlefish bones |
⁷: B. licheniformis followed by G. oxydans |
|
Table 3: Biological
process parameters.
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