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URL: https://pubmed.ncbi.nlm.nih.gov/34205202/

⇱ Isolation and Identification of Natural Colorant Producing Soil-Borne Aspergillus niger from Bangladesh and Extraction of the Pigment - PubMed

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Abstract

Natural colorants have been used in several ways throughout human history, such as in food, dyes, pharmaceuticals, cosmetics, and many other products. The study aimed to isolate the natural colorant-producing filamentous fungi Aspergillus niger from soil and extract pigments for its potential use specially for food production. Fourteen soil samples were collected from Madhupur National Park at Madhupur Upazila in the Mymensingh district, Bangladesh. The Aspergillus niger was isolated and identified from the soil samples by following conventional mycological methods (cultural and morphological characteristics), followed by confirmatory identification by a polymerase chain reaction (PCR) of conserved sequences of ITS1 ribosomal DNA using specific oligonucleotide primers. This was followed by genus- and species-specific primers targeting Aspergillus niger with an amplicon size of 521 and 310 bp, respectively. For pigment production, a mass culture of Aspergillus niger was conducted in Sabouraud dextrose broth in shaking conditions for seven days. The biomass was subjected to extraction of the pigments following an ethanol-based extraction method and concentrated using a rotary evaporator. Aspergillus niger could be isolated from three samples. The yield of extracted brown pigment from Aspergillus niger was 0.75% (w/v). Spectroscopic analysis of the pigments was carried out using a UV-VIS spectrophotometer. An in vivo experiment was conducted with mice to assess the toxicity of the pigments. From the colorimetric and sensory evaluations, pigment-supplemented products (cookies and lemon juice) were found to be more acceptable than the control products. This could be the first attempt to use Aspergillus niger extracted pigment from soil samples in food products in Bangladesh, but for successful food production, the food colorants must be approved by a responsible authority, e.g., the FDA or the BSTI. Moreover, fungal pigments could be used in the emerging fields of the food and textile industries in Bangladesh.

Keywords: Aspergillus niger; PCR; brown pigment; filamentous fungi; natural colorants.

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Conflict of interest statement

The authors do not have any conflict of interest.

Figures

👁 Figure 1
Figure 1
Pictorial view of sampling area ((A): map showing Bangladesh; (B): map showing Madhupur National Park; (C): sampling). Courtesy of (Accessed on 1 May 2021).
👁 Figure 2
Figure 2
A group of Swiss albino mice after 28 days of color feeding.
👁 Figure 3
Figure 3
Different unidentified soil-borne fungi (A–I) and a cultural characterization of Aspergillus niger (J).
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Figure 4
Microscopic morphology of Aspergillus niger viewed at 100×. The morphology of Aspergillus niger shows large, globose, dark brown conidial heads. Conidiophores were smooth walled, hyaline, or turning dark towards the vesicle.
👁 Figure 5
Figure 5
Identification of Aspergillus sp. and Aspergillus niger by polymerase chain reaction. (A) PCR identification of Aspergillus sp. by using genus-specific primers. Lane M- 100-bp DNA ladder, NC: negative control, PC: positive control, and lane 1–4 test samples. (B) PCR identification of Aspergillus niger by using specific primers. Lane M—1 Kb DNA ladder, NC: negative control, PC: positive control, and lane 1–3 test samples.
👁 Figure 6
Figure 6
Pigment extraction produced by Aspergillus niger. (A) Fermentation by Aspergillus niger; (B) filtrate after fermentation; (C) heat treatment of the filtrate; (D) final product found after rotary evaporation.
👁 Figure 7
Figure 7
UV-visible spectra of extracted fungal pigment; highest absorbance observed at 295 nm. (a) Negative control: distilled water; (b) 0.50 mL pigment diluted with 50 mL distilled water; (c) 1.0 mL pigment diluted with 50 mL distilled water.
👁 Figure 8
Figure 8
Absorbance values at different concentrations (0, 0.5 mL, 1.0 mL, and 2 mL) of diluted pigments in 50 mL of distilled water.
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Figure 9
Addition of extracted pigment with cookie dough and lemon juice.
👁 Figure 10
Figure 10
Comparison of L*, a*, b*, C*, and h between colored and colorless cookies and lemon juice, where, L*—lightness, read from 0 (black) to 100 (white); a*—(positive) red color, (negative) green color; b*—(positive) yellow color, (negative) blue color; C*—purity of the color; h—hue.

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