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Author(s): Tikendra Kumar Verma*1, Vijeyata Verma2, S.K. Jadhav3

Email(s): 1tiken03ymail.com, 2drvijeyataverma@gmail.com, 3jadhav9862@gmail.com

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    1Laxman Prasad Baidh Govt. Girls College, Bemetara, Chhattisgarh, India
    2Department of KriyaSharir, N.P.A. Govt. Ayurved College, Raipur, Chhattisgarh, India
    3School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India
    *Corresponding Author Email- tiken03@ymail.com

Published In:   Volume - 2,      Issue - 1,     Year - 2020


Cite this article:
Tikendra Kumar Verma, Vijeyata Verma, S.K. Jadhav (2020) Biosorption of Iron by pretreated fungal biomass of Aspergillus niger and Aspergillus flavus. NewBioWorld A Journal of Alumni Association of Biotechnology, 2(1):28-32.

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 NewBioWorld A Journal of Alumni Association of Biotechnology (2020) 2(1):28-32            

RESEARCH ARTICLE

Biosorption of Iron by pretreated fungal biomass of Aspergillus niger and Aspergillus flavus

Tikendra Kumar Verma1*, Vijeyata Verma2, S.K. Jadhav3

 

1Laxman Prasad Baidh Govt. Girls College, Bemetara, Chhattisgarh, India

2Department of KriyaSharir, N.P.A. Govt. Ayurved College, Raipur, Chhattisgarh, India

3School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India

Email: tiken03ymail.com; drvijeyataverma@gmail.com; jadhav9862@gmail.com

*Corresponding Author Email- tiken03@ymail.com

ARTICLE INFORMATION

 

ABSTRACT

Article history:

Received

25 November 2019

Received in revised form

18 January 2020

Accepted

20 January 2020

Keywords:

Iron;

pretreatment;

biosorption capacity; Aspergillus niger; Aspergillus flavus

 

 

The current biotechnology situation involves the discovery of a novel class of biosorbents with a high capacity for heavy metal removal. Iron (Fe2+) is one of the heavy metal, which is commonly found in earth’s crust and also in industrial process like a mining operation, steel and ferroalloy based industrial effluents. In the present study, The ability of Aspergillus niger and Aspergillus flavus fungal biomass to biosorption of iron  was examined in relation to the effects of physical pretreatment such as heat, autoclaving, freeze-drying, and chemical pretreatment such as sodium hydroxide, commercial laundry detergent, formaldehyde, acetic acid, and dimethyl sulfoxide. In comparison to live biomass, the maximum biosorption capacity of A. niger and A. flavus biomass after physical pretreatment, subjected to freeze drying, was 1.79 mg/0.1 g and 1.04 mg/0.1 g, respectively. After chemical pretreatment, subjected to acetic acid, it was 1.89 mg/0.1 g and 1.61 mg/0.1 g.

 


Introduction

The unceasing growth of industrialization and urbanization has increased the demand for natural resources. These kinds of human activities have also increased the level of pollution in the present environment day by day. One of the major pollution in present scenario is water pollution, which is due to the disposal of industrial effluent or domestic wastewater, many of which contains high levels of toxic substances including heavy metals. The metals such as Fe, Cu, Zn, Mn, Pb, Hg, Cd and Cr etc. are widely distributed into earth’s surface. These metal species are mobile in nature, which are released into the environment by technological activities of human incline to keep on completely accumulating throughout the food chain, ecosystem, humans and animals (Volesky and Holan, 1995).Iron is one of the heavy metal, which is widely used in electroplating industries, steel and ferroalloy units etc. Almost all organisms and living cell require iron for the basic cellular process. However, excessive iron causes iron toxicity. In humans, vomiting, diarrhea and damage in the intestine are the symptoms of iron toxicity and it is also affecting the aquatic life through getting precipitated in the gills of fishes (Binupriya et al., 2006).These are a serious concern to seek a solution for the removal of toxic elements from living organisms and provide them a healthy life cycle.

DOI: 10.52228/NBW-JAAB.2020-2-1-6

“Biosorption may be simply defined as the removal of substances from solution by biological material” (Gadd, 2009). There are various types of a biological system like bacteria, yeast, algae and fungi etc. are already investigated to remove heavy metals from aqueous solutions according to Wang and Chen (2009). Apart from these, fungi have a great potential to remove metallic substances, due to their easy availability, inexpensive growth media and lower maintenance, relatively to other biomaterials (Viraraghavan and Srinivasan, 2011).Huang and Huang (1996) suggested that pretreatment of fungal biomass increased the metal biosorption capacity due to the removal of surface impurities which reveals the available sites for metal binding. The present study investigates the use of biomass of two fungal species known as Aspergillus nige rand Aspergillus flavus for the biosorption of iron. The effects of different physical and chemical pretreatment of above fungal species on biosorption of iron (Fe2+) were investigated.

Methods and material

Preparation of biomass and pretreatment

The effluent from the steel industries was use for isolation of fungi. The fungi were isolated by serial dilution technique on Potato Dextrose Agar (PDA) and pure isolates were identified as Aspergillus niger and Aspergillus flavus on the basis of morphological and microscopic characteristics. These fungal cultures were routinely maintained on PDA. A liquid medium (YPG) (Yan and Viraraghavan, 2000) with pH value adjusted to 4.5 was prepared, which contains the following: yeast 3 gℓ-1, peptone 10 gℓ-1 and glucose 20 gℓ-1 for the production of fungal biomass. The culture was grown at 27°C in the medium in conical flasks kept on a rotary shaking incubator at 100 rpm for 7days. All culture works were done in the sterile condition. Fungal biomass was harvested by filtering the growth media through Whatman No. 1 filter paper for biosorption studies.

The harvested biomass washed with a generous amount of distilled water properly. The live biomass thus obtained was referred as Type A. After that, 15 g of Type A was pretreated with different physical and chemical methods as follow:

Physical treatment

·         Only dried at 60°C for 15h in a drying oven (Type B).

·         Autoclaved for 30 min. at 121°C, 15 psi (Type C).

·         Freeze-dried using liquid nitrogen to grind it (Type D).

Chemical treatment

·         250 ml of 0.5 N Sodium Hydroxide Solution (Type E) was boiled for 15 minutes.

·         250 ml of water containing 1.25 g of commercial laundry detergent (Type F) was boiled for 15 minutes.

·         250 ml of formaldehyde solution (Type G), 15% (vol/vol), boiled for 15 minutes.

·         200 ml of 10% (vol/vol) acetic acid solution (Type H) were boiled for 15 minutes.

·         200 ml of 50% (vol/vol) dimethyl sulfoxide solution (Type I) were boiled for 15 minutes.

After each pretreatment with chemicals, the biomass was washed with a generous amount of distilled water and then dried at 60°C in a drying oven for 15h. Sodium hydroxide pretreated biomass was washed with distilled water up to the pH of the wash solution was near in neutral range (pH 6.8-7.2). Dried biomass was ground using mortar pestle.

Biosorption studies

Biosorption experiment carried out using iron (Fe2+) containing solution (in the form of Ammonium ferrous sulfate [(NH4)2Fe(SO4)2.6H2O]) prepared in distilled water and the initial metal ion concentration was approximately 5 mgℓ-1 in the solution. Each type of pretreated biomass (0.1g) was added to 50 mℓ of iron solution at pH 5.5. On a rotary shaker, the reaction mixture was stirred at a speed of 100 rpm. After 15 hours of contact, the reaction mixture was filtered using Whatman No. 1 filter paper to separate the biomass, and the iron content of the filtrate solution was assessed. Iron concentration was measured using an Iron test kit (Iron Test 0.005-5.00 mgℓ-1, Merck, Germany) and its absorbance level was measured by Spectroquant NOVA 60 (Merck, Germany). Biosorption experiment was performed in duplicates and average values were used in the analysis. The following equation was used to compute biosorption capacity, or the amount of metal ion (mg) that was biosorbed into each gramme (dry weight) of biomass:

Q = (Ci – Cf /m) V

Where, Q = amount of metal ion bisorbed per g of biomass (mg); Ci= initial metal ion concentration (mgℓ-1); Cf= final metal ion concentration (mgℓ-1); m = mass of biomass in the reaction mixture (g); V = volume of the reaction mixture (ℓ), (Kapoor and Viraraghavan, 1997).

Results

The live biomass of A. niger (0.91mg/0.1g) showed the highest biosorption capacity for Fe2+ ions in comparison with A. flavus (0.84 mg/0.1g). In comparison to live biomass, the biosorption of iron either increased or reduced depending on the pretreatment method. The results related to Fe2+ biosorption by live and pretreated fungal biomass of A. niger are presented in Figure 1. Pretreatment of live biomass using all method heat, autoclaved, freeze-dried, NaOH, detergent, formaldehyde, acetic acid and DMSO resulted in an enhancement of iron biosorption by A. niger compared to live biomass (from 0.91to 1.34-1.89 mg/0.1g).

The results of pretreatment of A. flavus related to biosorption of iron were shown in Figure 2. Pretreatment of biomass using heat, freeze-dried, formaldehyde, acetic acid and DMSO resulted in an enhancement of iron biosorption in comparison with live biomass (from 0.84 to 0.97-1.61mg/0.1g) while autoclaved, NaOH and detergent were significantly reduced the iron biosorption by A. flavus in comparison with live biomass (from 0.84 to 0.37-0.47 mg/0.1g).The actual enhancement (%) of biosorption of iron by different physically and chemically pretreated biomass of A. niger and A. flavus was shown in Table 1, in comparison with live biomass of both fungi, respectively.


Table 1: Actual enhancement (%) of biosorption of iron by pretreated fungal biomass in comparison with live biomass

Fungal biomass

Pretreatment Method

B

C

D

E

F

G

H

I

A. niger

55.05

53.39

97.43

47.34

91.38

93.03

108.44

80.37

A. flavus

15.48

-44.05

23.81

-55.95

-48.81

83.33

91.67

64.29

 

Figure1: Fe2+biosorption by live (Type A) and pretreated biomass (TypeB-I) of Aspergillus niger

Figure 2: Fe2+biosorption by live (Type A) and pretreated biomass (Type B-I) of Aspergillus flavus


Discussion

It was observed that Q values obtained from all the physically treated biomasses were high in comparison to live biomass for both fungi A. niger and A. flavus, except autoclaved biomass (Type C) of A. flavus obtained low value. Freeze dried biomasses (Type D) showed maximum improvement on iron biosorption for both fungi A. niger and A. flavus, from 0.91 to 1.79 mg/0.1g and from 0.84 to 1.04mg/0.1g, respectively. During the process of freeze drying, several microscopic pores would be formed inside the fungal biomass due to the formation of ice crystals that channel which provide more surface area for biosorption. Similar results found by Das et al. (2007) for cadmium sorption. Cabuk et al. (2005) suggested that the drying and then grinding of fungal biomass expose more sites for metal binding, therefore, the probability of seizing metal ions could be increased. The finding showed that heat and autoclaved biomass of A. niger increased the biosorption capacity rather than biosorption capacity of A. flavus which reduced in autoclaved biomass comparison with live biomass, respectively. Yan and Viraraghavan (2000) also reported that pretreated with autoclaved biomass of Mucor rouxii reduced the biosorption capacity of heavy metal due to the lake of intracellular uptake. In the same way, Kapoor and Viraraghavan (1998) reported that A. niger pretreated with autoclave decreased the biosorption of lead, cadmium, copper and nickel.

The Q values obtained from all the chemically treated biomasses were maximum in comparison with live biomass for both fungi A. niger and A. flavus, except NaOH and detergent treated biomass (Type E and F) of A. flavus obtained low value. Pretreatment with acetic acid (Type H) significantly increased biosorption of iron for both fungi A. niger and A. flavus from 0.91 to 1.89 mg/0.1g and from 0.84 to 1.61mg/0.1g, respectively. Kapoor and Viraraghavan (1998) also reported that A. niger pretreated with acetic acid enhanced the biosorption capacity of lead and copper. In the same way, Cabuk et al. (2005) were obtained that acetic acid pretreatment increased biosorption of lead by fungal biomass of Metarrhizium anisopliae var anisopliae and Penicillium verrucosum. Pretreatment of live biomass using formaldehyde (Type G) and DMSO (Type I) also increased biosorption of iron by A.  niger (from 0.91 to 1.64-1.75 mg/0.1g) and A. flavus (0.84 to 1.38-1.54 mg/0.1g).Similar improvement in biosorption was reported by Cabuk et al. (2005) for the lead, Kapoor and Viraraghavan (1998)for lead, cadmium and copper and Ilhan et al.(2004)for copper metal ion using formaldehyde pretreated biomass. Pretreatment with DMSO of fungal biomass was also suggested by Kiran et al. (2005) for lead and copper, Cabuk et al. (2005) for lead and Kapoor and Viraraghavan (1998) for lead, cadmium and copper to the improvement of biosorption capacity.

Pretreatment of fungal biomass using NaOH (Type E) and detergent (Type F) were also increased biosorption capacity of A. niger (from 0.91 to 1.34-1.74 mg/0.1g)in comparison with live biomass. In contrast, pretreatment using NaOH and detergent significantly reduced biosorption capacity of A. flavus in comparison with live biomass (from 0.84 to 0.43-0.37mg/0.1g). Kapoor and Viraraghavan (1998) showed that similar results to enhance the biosorption capacity for lead, cadmium and copper using NaOH and detergent except for nickel biosorption which reduced in comparison with live biomass.

Yan and Viraraghavan (2000) suggested that alkali treatment such as NaOH with boiling caused heavy loss of biomass, therefore, the significant reduction of biosorption capacity of metal ions in comparison with NaOH with autoclaved biomass. Most of the detergent also contains alkalis as an ingredient; therefore, pretreatment with alkaline detergent resulted in an enhancement of biosorption of metal ions. The fungal species belong to different genera or same genera have different chemical characteristics in their cell wall composition, which is one of the reasons why metal biosorption capacities are different from each other (Ilhan et al., 2004, Cabuk et al., 2005).

Conclusion

Biosorption capacity of dead fungal biomass may be more useful than the use of live biomass, depending upon the pretreatment method applied. The findings suggest that iron can be removed from industrial effluents using biomasses of A. niger and A. flavus that have undergone physical and chemical pretreatment procedures. Utilizing A. niger that has already been treated with acetic acid may be useful if the removal of iron (Fe2+) is necessary. It is also necessary to get the knowledge about the chemical composition of fungal cell walls, which suits the best combination of particular metal ions, pretreatment methods and other condition.  Furthermore, details study should be required to understand causes of enhancement or reduction in biosorption capacity of microbial biomass.

Acknowledgment

The authors gratefully acknowledge to the funding provided by DST, New Delhi in the form of DST-FIST (Sl. No. 270 for tenure of 2013-18) and also acknowledge PRSU Raipur, for providing financial assistance to T. K. Verma in the form of University Research Fellowship.

References

Binupriya AR, Sathishkumar M, Swaminathan K, Jeong ES, Yun SE and Pattabi S(2006) Biosorption of metal ions from aqueous solution and electroplating industry wastewater by Aspergillus japonicus: Phytotoxicity studies. Bull. Environ.Contam.Toxicol.77 219–227.

Cabuk A, Ilhan S, Cansu F and Caliskan F (2005) Pb2+ Biosorption by pretreated fungal biomass. Turkish J. Biol.29 23–28.

Das N, Charumathi D andVimala R (2007) Effect of pretreatment on Cd2+ biosorption by mycelial biomass of Pleurotus florida. J. Biotechnol.6(22) 2555–2558.

GaddGM (2009) Biosorption: critical review of scientific rationale, environmental importance and significance for pollution treatment. J. Chem. Technol. Biotechnol.84 13–28.

Huang C and Huang CP (1996) Application of Aspergillus oryzae and Rhizopus oryzae for Cu (II) removal.Water Res.30(9) 1985-1990.

Ilhan SI, Cabuk A, Filik C and Caliskan F (2004) Effect of Pretreatment on Biosorption of Heavy Metals By Fungal Biomass. Trakya Univ. J. Sci.5(1) 11–17.

KapoorA and Viraraghavan T (1997) Heavy Metal Biosorption Sites in Aspergillus niger.Bioresour. Technol.61 221–227.

KapoorA and ViraraghavanT (1998) Biosorption of heavy metals on Aspergillus niger: Effect of pretreatment.Bioresour. Technol.63 109–113.

Kiran I, Akar T and Tunali S (2005) Biosorption of Pb(II) and Cu(II) from aqueous solutions by pretreated biomass of Neurospora crassa. Process Biochem.40 3550–3558.

Viraraghavan T and Srinivasan A (2011) Fungal biosorption and biosorbents. In: Kotrba P, Mackova M and MacekT (eds.) Microbial Biosorption of Metals. Springer Science, Dordrecht. pp. 143–158.

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