Akram Zamani

Mucor indicus: biology and industrial application perspectives: a review.

Mucor indicus, one of the most important strains of zygomycetes fungi, has been the subject of several studies since a couple of hundred years ago. This fungus, regarded as a non-pathogenic dimorphic microorganism, is used for production of several beers and foods. Morphology of the fungus can be manipulated and well controlled by changing a number of parameters. Furthermore, M. indicus can grow on a variety of substrates including lignocellulosic hydrolysates which are mixtures of hexoses, pentoses, and different severe fermentation inhibitors. Indeed, high yield ethanol production is among the most important features of this strain. Presence of considerable amounts of chitosan in the cell wall is another important aspect of the fungus. Besides production of ethanol and chitosan, the biomass of this fungus has shown a great potential to be used as a rich nutritional source, e.g. fish feed. The fungus is also among the oleaginous fungi and produces high amounts of polyunsaturated fatty acids particularly γ-linolenic acid. Furthermore, the biomass autolysate has a high potential for yeast extract replacement in fermentation by the fungus. Additionally, the strain has shown promising results in heavy metal removal from wastewaters. This review discusses different aspects of biology and industrial application perspectives of M. indicus. Furthermore, open areas for the future basic and applied levels of research are also presented.

Determination of Glucosamine and N-Acetyl Glucosamine in Fungal Cell Walls

A new method was developed to determine glucosamine (GlcN) and N-acetyl glucosamine (GlcNAc) in materials containing chitin and chitosan, such as fungal cell walls. It is based on two steps of hydrolysis with (i) concentrated sulfuric acid at low temperature and (ii) dilute sulfuric acid at high temperature, followed by one-step degradation with nitrous acid. In this process, chitin and chitosan are converted into anhydromannose and acetic acid. Anhydromannose represents the sum of GlcN and GlcNAc, whereas acetic acid is a marker for GlcNAc only. The method showed recovery of 90.1% of chitin and 85.7-92.4% of chitosan from commercial preparations. Furthermore, alkali insoluble material (AIM) from biomass of three strains of zygomycetes, Rhizopus oryzae, Mucor indicus, and Rhizomucor pusillus, was analyzed by this method. The glucosamine contents of AIM from R. oryzae and M. indicus were almost constant (41.7 +/- 2.2% and 42.0 +/- 1.7%, respectively), while in R. pusillus, it decreased from 40.0 to 30.0% during cultivation from 1 to 6 days. The GlcNAc content of AIM from R. oryzae and R. pusillus increased from 24.9 to 31.0% and from 36.3 to 50.8%, respectively, in 6 days, while it remained almost constant during the cultivation of M. indicus (23.5 +/- 0.8%).

Effect of carboxymethylation conditions on the water-binding capacity of chitosan-based superabsorbents

A superabsorbent polymer (SAP) from chitosan was provided via carboxymethylation of chitosan, followed by cross-linking with glutaraldehyde and freeze-drying. This work was focused on an investigation of the effects of monochloroacetic acid (MCAA), sodium hydroxide, and reaction time on preparation of carboxymethyl chitosan (CMCS). The CMCS products were characterized using FTIR spectroscopy, and their degrees of substitution (DS) were measured using conductimetry and FTIR analysis. The highest DS value was obtained when the carboxymethylation reaction was carried out using 1.75g MCAA and 1.75g NaOH per g of chitosan in 4h. The water solubilities of the CMCS products at various pHs were also evaluated, and the results indicated a significant impact of the reaction parameters on the solubility of CMCS. The CMCSs with the highest DS value resulted in SAPs having the highest water-binding capacity (WBC). The WBC of the best SAP measured after 10min exposure in distilled water, 0.9% NaCl solution, synthetic urine, and artificial blood was 104, 33, 30, and 57g/g, respectively. The WBC of this SAP at pH 2-9 passed a maximum at pH 6.

Temperature Shifts for Extraction and Purification of Zygomycetes Chitosan with Dilute Sulfuric Acid

The temperature-dependent hydrolysis and solubility of chitosan in sulfuric acid solutions offer the possibility for chitosan extraction from zygomycetes mycelia and separation from other cellular ingredients with high purity and high recovery. In this study, Rhizomucor pusillus biomass was initially extracted with 0.5 M NaOH at 120 °C for 20 min, leaving an alkali insoluble material (AIM) rich in chitosan. Then, the AIM was subjected to two steps treatment with 72 mM sulfuric acid at (i) room temperature for 10 min followed by (ii) 120 °C for 45 min. During the first step, phosphate of the AIM was released into the acid solution and separated from the chitosan-rich residue by centrifugation. In the second step, the residual AIM was re-suspended in fresh 72 mM sulfuric acid, heated at 120 °C and hot filtered, whereby chitosan was extracted and separated from the hot alkali and acid insoluble material (HAAIM). The chitosan was recovered from the acid solution by precipitation at lowered temperature and raised pH to 8–10. The treatment resulted in 0.34 g chitosan and 0.16 g HAAIM from each gram AIM. At the start, the AIM contained at least 17% phosphate, whereas after the purification, the corresponding phosphate content of the obtained chitosan was just 1%. The purity of this chitosan was higher than 83%. The AIM subjected directly to the treatment with hot sulfuric acid (at 120 °C for 45 min) resulted in a chitosan with a phosphate impurity of 18.5%.

Enhanced Solid-State Biogas Production from Lignocellulosic Biomass by Organosolv Pretreatment

Organosolv pretreatment was used to improve solid-state anaerobic digestion (SSAD) for methane production from three different lignocellulosic substrates (hardwood elm, softwood pine, and agricultural waste rice straw). Pretreatments were conducted at 150 and 180°C for 30 and 60 min using 75% ethanol solution as an organic solvent with addition of sulfuric acid as a catalyst. The statistical analyses showed that pretreatment temperature was the significant factor affecting methane production. Optimum temperature was 180°C for elmwood while it was 150°C for both pinewood and rice straw. Maximum methane production was 152.7, 93.7, and 71.4 liter per kg carbohydrates (CH), which showed up to 32, 73, and 84% enhancement for rice straw, elmwood, and pinewood, respectively, compared to those from the untreated substrates. An inverse relationship between the total methane yield and the lignin content of the substrates was observed. Kinetic analysis of the methane production showed that the process followed a first-order model for all untreated and pretreated lignocelluloses.

Characterization of Nizimuddinia zanardini macroalgae biomass composition and its potential for biofuel production

Nizimuddinia zanardini macroalgae, harvested from Persian Gulf, was chemically characterized and employed for the production of ethanol, seaweed extract, alginic acid, and biogas. In order to improve the products yields, the biomass was pretreated with dilute sulfuric acid and hot water. The pretreated and untreated biomasses were subjected to enzymatic hydrolysis by cellulase (15FPU/g) and β-glucosidase (30IU/g). Hydrolysis yield of glucan was 29.8, 82.5, and 72.7g/kg for the untreated, hot-water pretreated, and acid pretreated biomass, respectively. Anaerobic fermentation of hydrolysates by Saccharomycescerevisiae resulted in the maximum ethanol yield of 34.6g/kg of the dried biomass. A seaweed extract containing mannitol and a solid residue containing alginic acid were recovered as the main byproducts of the ethanol production. On the other hand, the biogas yield from the biomass was increased from 170 to 200m(3) per ton of dried algae biomass by hot water pretreatment.

Optimization of xanthan gum production using cheese whey and response surface methodology

Cheese whey lactose was used as a carbon source for xanthan gum production with Xanthomonas campestris and Xanthomonas pelargonii. Proteins were precipitated and removed from whey prior to fermentation. Box-Behnken response surface methodology was used for optimization of the carbon, magnesium, and phosphate source concentrations in the culture medium to maximize xanthan gum production. After 48 h of fermentation using X. campestris, the highest xanthan concentration (16.4 g/L) was achieved at 65.2 g/L of cheese whey (39.1 g/L of lactose), 14.8 g/L of phosphate (K H2PO4), and 1.1 g/L of magnesium (MgSO4·7H2O). The corresponding optimum cheese whey, phosphate, and magnesium concentrations in cultures of X. pelargonii were 80.0, 6.7, and 0.8 g/L, respectively, which resulted in a xanthan production of 12.8 g/L. The xanthan gum yield (g of xanthan/g of lactose) was 0.42 for X. campestris and 0.27 for X. pelargonii.

Determination of glucosamine in fungal cell walls by high-performance liquid chromatography (HPLC).

Glucosamine (GlcN) is a major and valuable component in the cell wall of zygomycetes fungi. In this study, a time independent and accurate method was developed for the determination of GlcN. In this method, the cell wall was treated via a two-stage sulfuric acid process, and chitin and chitosan were fully deacetylated, partially depolymerized, and converted to GlcN oligosaccharides. Then, the oligosaccharides were deaminated to 2,5-anhydromannose using nitrous acid. Finally, 2,5-anhydromannose was analyzed by high performance liquid chromatography (HPLC). The determinations of pure GlcN solutions were stable at least for 10 days, while those of the conventional colorimetric method were not stable for more than one hour. The alkali insoluble material (AIM) of biomass of purely yeast-like, mostly yeast-like, and filamentous forms of the fungus Mucor indicus was analyzed by the developed method. The respective GlcN content of AIM of the fungus was 0.232, 0.204, and 0.458 (g/g).

Production of xanthan gum by free and immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii.

Production of xanthan gum using immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii grown on glucose or hydrolyzed starch as carbon sources was investigated. Calcium alginate (CA) and calcium alginate-polyvinyl alcohol-boric acid (CA-PVA) beads were used for the immobilization of cells. Xanthan titers of 8.2 and 9.2g/L were obtained for X. campestris cells immobilized in CA-PVA beads using glucose and hydrolyzed starch, respectively, whereas those for X. pelargonii were 8 and 7.9 g/L, respectively. Immobilized cells in CA-PVA beads were successfully employed in three consecutive cycles for xanthan production without any noticeable degradation of the beads whereas the CA beads were broken after the first cycle. The results of this study suggested that immobilized cells are advantageous over the free cells for xanthan production. Also it was shown that the cells immobilized in CA-PVA beads are more efficient than cells immobilized in CA beads for xanthan production.

A sulfuric–lactic acid process for efficient purification of fungal chitosan with intact molecular weight

The most recent method of fungal chitosan purification, i.e., two steps of dilute sulfuric acid treatment, pretreatment of cell wall at room temperature for phosphate removal and extraction of chitosan from the phosphate free cell wall at high temperature, significantly reduces the chitosan molecular weight. This study was aimed at improvement of this method. In the pretreatment step, to choose the best conditions, cell wall of Rhizopus oryzae, containing 9% phosphate, 10% glucosamine, and 21% N-acetyl glucosamine, was treated with sulfuric, lactic, acetic, nitric, or hydrochloric acid, at room temperature. Sulfuric acid showed the best performance in phosphate removal (90%) and cell wall recovery (89%). To avoid depolymerisation of chitosan, hot sulfuric acid extraction was replaced with lactic acid treatment at room temperature, and a pure fungal chitosan was obtained (0.12 g/g cell wall). Similar pretreatment and extraction processes were conducted on pure shrimp chitosan and resulted in a chitosan recovery of higher than 87% while the reduction of chitosan viscosity was less than 15%. Therefore, the sulfuric-lactic acid method purified the fungal chitosan without significant molecular weight manipulation.