Resources, Environment and Sustainability 5 (2021) 100030

Contents lists available at ScienceDirect

Resources, Environment and Sustainability

journal homepage: www.elsevier.com/locate/resenv

Evaluation of Aspergillus fumigatus NTCC1222 as a source of enzymes for

detergent industry
Shalini Singh ∗, Jyoti Mangla, Sanamdeep Singh
School of Bioengineering and Biosciences, Lovely Professional University, Punjab, 144411, India

ARTICLE INFO ABSTRACT

Keywords: Enzymes are playing crucial roles in improving the product quality in a number of conventional chemical based
Aspergillus fumigatus NTCC1222 industries including detergent industry. Amylases are widely used in detergent industry as starch is a major
Amylase component of various types of stains and amylases degrade starch. Therefore, amylases and other detergent
Detergent formulations
enzymes can replace some chemicals used for starch degradation in detergent industries. In the present study,
Detergent industry
the compatibility of the amylase produced by wild-type and mutated strains of Aspergillus fumigatus NTCC1222
Water quality
with various commercially available detergent formulations was checked. The quality of water used also play
an important role in determining the efficacy of enzymes in detergent formulations. The influence of hardness
of water (artificial hard water) used for preparing the detergent formulations on enzyme activity and stability,
in comparison to tap water, under different incubation periods was thus also studied. The enzyme was highly
compatible with all the detergent formulations evaluated during the study for at least one of the incubation
periods tested, indicating that the enzyme could be very well used in detergent formulations. The enzyme was
active even in hardwater though a loss in its activity was observed as compared to distilled water.

1. Introduction
In today’s world there is a great need to replace chemicals with
environment friendly compounds. The search for alternative ‘environ-
ment friendly’ technologies has made the biotechnological approach
meaningful in the present context where, microorganisms and their
products are being used in place of chemicals. As with other industrial
applications, cost effectiveness is also one of the major concerns that
influence the successful use of biotechnological applications in indus-
tries (Balkan and Ertan, 2007). Enzymes, especially from microbial
sources, are the most sought after products to meet the expectations
of environment friendly technologies. These are rapidly being adopted
in many conventional chemical based industries like, leather, paper,
detergent, textile, etc. (Akpan et al., 1999; Anto et al., 2006; Pandey
et al., 2000; Khambhaty, 2020; Singh et al., 2010; Gurkok, 2019;
Singh, 2016). Along with industrially important enzymes like, cellu-
lases, lipases, hemicellulases, proteases, etc., amylases have exhibited
great potential a wide number of industrial processes (de Souza and
de Oliveira Magalhães, 2010; Sindhu et al., 2011), including food, tex-
tile, paper and detergent industry (de Souza and de Oliveira Magalhães,
2010). The use of enzymes in detergent formulations is now common
in developed countries. Though the detergent industry is one of the
largest market for enzymes, both volume wise as well as value wise (de
Souza and de Oliveira Magalhães, 2010), yet details of the enzymes
used and the ways in which they are used, have rarely been published.
Clothes need to be free from dirt (appearing in different forms and
includes proteins, starches and lipids). Using detergents in water at
high temperatures and with vigorous mixing, it is possible to remove
most types of dirt but the cost of heating the water is high and lengthy
mixing or beating will shorten the life of clothing and other materials.
The use of enzymes allows lower temperatures to be employed and
shorter periods of agitation are needed, often after a preliminary period
of soaking. The use of enzymes in detergents formulations enhances
the detergents ability to remove tough stains and making the detergent
environmentally safe. Amylases are one of the most important enzymes
used in the formulation of enzymatic detergent with, 90% of all liquid
detergents containing these enzymes (Sindhu et al., 2011).
Fungal amylases, as is true for other fungal enzymes hold much
more importance as compared to bacterial ones for obvious and clearly
established reasons (Sindhu et al., 2011). Also, the cultivation of
filamentous fungi for extracellular enzyme production under solid
state fermentation conditions further improve their industrial appli-
cations for their specific advantages over liquid state fermentation
conditions (Singh et al., 2009) Reports on application of enzymes in
detergent industry have been reported (Gurkok, 2019; Khajuria and
Singh, 2020) using few microbial cultures like, Aspergillus niger (Mi-
tidieri et al., 2006), Penicillium janthinellum NCIM 4960 (Sindhu et al.,
2011), Bacillus sp. (Borchert et al., 1995; Rodríguez et al., 2006; de
Carvalho et al., 2008).

∗ Corresponding author.
E-mail address: shalinisingh.iit@gmail.com (S. Singh).

https://doi.org/10.1016/j.resenv.2021.100030
Received 1 January 2021; Received in revised form 27 May 2021; Accepted 25 June 2021
Available online 9 July 2021
2666-9161/© 2021 The Authors. Published by Elsevier B.V. on behalf of Lishui Institute of Ecology and Environment, Nanjing University. This is an open
access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
S. Singh, J. Mangla and S. Singh Resources, Environment and Sustainability 5 (2021) 100030

Table 1
Stability of amylase of wild type strain of A. fumigatus NTCC1222 in commercial detergent formulations prepared in different quality of water.
Type of water Sample Amylase activity (U/mL)/% increase/decrease in amylase activity for different
detergent formulations compared to corresponding control (+∕−)
20 mins 40 mins 60 mins 80 mins 120 mins
Distilled Crude enzyme w/o 337.30 ± 1.04 338.90 ± 0.67 344.90 ± 0.82 318.30 ± 0.45 288.60 ± 1.21
detergent (control) U/ml U/ml U/ml U/ml U/ml
E with D1 +02.80 +22.60 +30.50 +31.90 +24.90
E with D2 +08.10 +16.70 +29.20 +24.90 +23.20
E with D3 +02.30 +23.10 +10.90 +08.02 −09.60
E with D4 +07.50 +08.10 +15.06 +30.20 +13.60
E with D5 +05.35 +08.50 +23.20 +14.60 +06.30
E with D6 +01.10 +02.17 −08.80 −44.20 −62.56
E with D7 +29.90 −32.10 −38.00 −64.20 −76.78
Tap Crude enzyme w/o 328.20 ± 1.09 331.30 ± 1.22 342.30 ± 0.98 333.70 ± 0.45 325.20 ± 0.82
detergent (control) U/ml U/ml U/ml U/ml U/ml
E with D1 +27.70 +28.10 +30.10 +32.80 +33.10
E with D2 +08.80 +31.30 +37.70 +42.90 +48.80
E with D3 +04.70 +11.20 +37.70 +44.70 +43.40
E with D4 +16.10 +10.20 +00.77 −03.80 −09.84
E with D5 +04.70 +17.05 +15.80 +05.98 −03.80
E with D6 +00.20 +00.10 −08.00 −14.50 −13.20
E with D7 +06.20 +03.50 +02.06 −05.57 −04.75
Crude enzyme w/o 177.4 ± 0.56 189.30 ± 0.78 236.20 ± 0.77 171.0 ± 1.11 152.50 ± 1.02
detergent (control) U/ml U/ml U/ml U/ml U/ml
E with D1 +05.93 +16.64 +20.50 +27.60 +44.20
HARD
E with D2 +32.10 +32.40 +31.80 +06.08 +06.20
E with D3 −08.90 +13.40 +16.20 +17.90 +02.30
E with D4 +31.60 +20.60 +22.80 +05.13 +01.60
E with D5 +02.30 +07.30 +16.90 +07.80 +09.80
E with D6 +01.50 +04.00 +22.30 −35.60 −58.90
E with D7 +11.60 +15.40 +32.90 −15.60 −26.40
Assay conditions:
Temperature, ◦ C : 55
Time, minutes : 60

*+∕−: % increase/decrease in amylase activity for different detergent formulations compared to corresponding control
**E: Enzyme
***D: Detergent formulations
****U/mL: Amylase activity in Units/ml
*****±: standard deviation from the mean.

To the best of our knowledge, crude amylase preparations of As-
pergillus fumigatus for biocompatibility evaluations in detergent for-
mulations have not been extensively reported and the simultaneous
influence of detergent formulations as well as quality of water used
for preparing detergent solutions is further rarely reported. Keeping
in view the significance of cheaply produced and effective microbial
enzymes for detergent industry, and under-exploration of Aspergillus
fumigatus’ amylases for detergent formulations, the current study aims
to draw a comparison of compatibility of crude amylase preparation
obtained from wild-type and mutated strains of A. fumigatus NTCC 1222
with various commercially detergents available in the Indian market.
A. fumigatus NTCC 1222, as a promising source of amylases, which
can be further explored for industrial applications, has already been
reported (Singh et al., 2014, 2013a,b,?,d, 2015, 2016) and the current
investigations further explore its use in another important significantly
chemical based detergent industries.
2. Materials and methods
Starch soluble and 3, 5-Dinitrosalicylic acid (DNS) reagent were of
analytical grade and procured from Himedia Pvt. Ltd. India. All other
chemical reagents and nutrient culture media used were procured from
Titan Biotech Pvt. Ltd. India and Lobachemie Pvt Ltd., India. Lignocel-
lulosic solid substrates were procured from local market of Kapurthala,
Punjab, India. Detergents were purchased from the local market (The
commercial names of the detergents have not been disclosed in the
current study).
Wild-type and chemically mutated strains of Aspergillus fumigatus
NTCC 1222 was used in the present study (Singh et al., 2014, 2016).
For short duration, the fungal strains were subcultured on potato
dextrose agar (PDA) medium, incubated at 37 ◦ C for 3 days and
subsequently stored at 4 ◦ C. For long term preservation, the fungal
culture was stored as glycerol stocks in the form of spore suspension
at −20 ◦ C.
The test strain was subjected to optimized conditions of solid state
fermentation (SSF) including, incubation period (6 days), pH (6), tem-
perature (37 ◦ C), beef extract as nitrogen source, wheat bran as the
substrate and wheat bran to moistening agent ratio of 1:3 (Singh et al.,
2013b). The crude enzyme was harvested (Singh et al., 2009), and the
amylase activity was determined in terms of reducing sugars released
and subsequently reported as U/mL (Miller, 1959). The crude enzyme
preparation obtained from each respectively, was designated as Enzyme
A and B for future reference.
The Enzyme A and Enzyme B, obtained respectively from wild-type
and mutated strains of A. fumigatus NTCC 1222, produced as above,
were used in the current study.
Distilled water (produced as per standard distillation process under
laboratory conditions), Tap water (Supplied from Municipal Corpora-
tion, Jalandhar, Punjab, India) and artificial hard water (addition of
150 mg/l of magnesium carbonate to distilled water) were used in the
present study.
For potential commercial exploitation of the enzyme, the enzyme
A and B were analyzed for its compatibility with commercial 7 dif-
ferent commercially available detergent formulations. These detergent
formulations were designated as D1, 2, 3, 4, 5, 6, and 7. The effect
of hardness of water on compatibility of amylase preparation with
detergent formulation was also evaluated so as to determine the in-
fluence of water composition on the activity of the enzyme as well.

2
S. Singh, J. Mangla and S. Singh Resources, Environment and Sustainability 5 (2021) 100030

Table 2
Stability of amylase of chemically mutated strain of A. fumigatus NTCC1222 in commercial detergent formulations prepared in different quality of water.
Type of water Sample +∕−: % increase/decrease in amylase activity for different detergent formulations
compared to corresponding control
20 mins 40 mins 60 mins 80 mins 120 mins
Distilled Crude enzyme w/o 699.40 ± 0.09 747.20 ± 0.77 814.50 ± 0.56 761.50 ± 1.05 707.40 ± 1.23
detergent (control) U/ml U/ml U/ml U/ml U/ml
E with D1 +18.05 +12.70 −04.90 −19.60 −35.80
E with D2 +19.40 +14.40 +08.70 +08.50 −35.60
E with D3 +16.80 +10.40 +03.20 +02.50 −44.90
E with D4 +05.50 −05.50 −11.27 −17.20 −48.07
E with D5 +05.90 +00.40 −05.30 −15.90 −22.50
E with D6 +01.60 −08.80 −13.73 −25.30 −56.60
E with D7 +13.40 −07.10 −15.10 −19.90 −17.10
Tap Crude enzyme w/o 725.00 ± 0.15 711.20±U/ml 741.20±U/ml 772.50±U/ml 801.30±U/ml
detergent (control) U/ml
E with D1 +07.90 +01.60 +01.70 −09.80 −11.20
E with D2 +04.90 +06.90 +12.60 +01.40 +00.80
E with D3 +03.06 +05.57 +09.52 −07.40 −08.50
E with D4 +03.73 +05.50 +07.70 +25.40 +30.60
E with D5 +08.00 +04.90 +02.50 −14.80 −20.60
E with D6 −01.50 −11.30 −16.20 −46.40 −50.40
E with D7 −01.70 −03.60 −06.40 −09.15 −05.10
HARD Crude enzyme w/o 320.20 ± 0.77 387.80 ± 0.56 478.60 ± 1.04 357.40 ± 2.01 334.00 ± 1.95
detergent (control) U/ml U/ml U/ml U/ml U/ml
E with D1 +41.80 +58.20 +22.10 +13.60 +10.90
E with D2 +38.90 +36.80 +32.40 +32.06 −02.40
E with D3 +06.40 +25.08 +19.60 +11.30 +09.60
E with D4 +32.50 +16.90 +15.30 +11.10 −15.90
E with D5 +38.10 +04.40 −19.10 −24.50 −29.10
E with D6 +30.80 +03.35 −29.70 −34.60 −60.20
E with D7 +35.10 +20.07 −12.10 −47.60 −62.80
Assay conditions:
Temperature, ◦ C : 55
Time, minutes : 60

*+∕−: % increase/decrease in amylase activity for different detergent formulations compared to corresponding control
**E: Enzyme
***D: Detergent formulations
****U/ml: Amylase activity in Units/ml
*****±: standard deviation from the mean.

The detergents, D1 to D7, were dissolved in distilled, tap water and
hard water at a concentration of 7 mg/ml and the detergent solutions
so prepared were incubated at 100 ◦ C for 15 min to deactivate the
already present enzymes in detergent formulations, if any (Rodríguez
et al., 2006). 75 U/ml of crude amylase and 500 𝜇l of 1% (v/v)
starch were mixed with 400 𝜇l of inactivated detergent solutions. The
reaction mixture, so obtained, was incubated at 55 ◦ C for variable
incubation period (20, 40, 60, 80 and 120 min). After appropriate
incubation time, 3 ml DNS (Dinitrosalicylic acid) reagent was added to
each sample and kept in boiling water at 100 ◦ C for 10 min to stop
the reaction. Absorbance for each sample was measured at 540 nm
and amylase activity determined using glucose standard curve (Miller,
1959). Appropriate control (reaction mixture of crude enzyme and
starch incubated without detergent solution) was also assayed using
DNS test for comparison.
All experiments to check enzyme activity were carried out in tripli-
cates and the results were mean ± standard deviation (SD) of the values.
The findings have been reported in the current manuscript in ‘% in-
crease/decrease in amylase activity for different detergent formulations
compared to corresponding control’. For clarity, the enzyme values for
control have been given real time, with standard deviation, while for
the rest, ‘% increase/decrease in amylase activity has been given.
3. Results and discussion
The stability of enzyme was checked at different incubation time
(Table 1). It was found that the enzyme activity increased up to 60
mins of incubation period for all types of water samples used (344.90
U/ml for distilled water, 342.30 U/ml for tap water and 236.20 U/
ml for hard water. Thereafter, the enzyme activity decreased in all
the cases. In case of wild-type strain of A. fumigatus NTCC1222, the
enzyme activity was found to improve in the presence of all detergents
at least, one incubation period for all types of water samples used
(distilled, tap and hard water). In the presence of distilled water,
enzyme activity showed highest improvement of 31.10% at 80 mins
of incubation period for detergent D1. Only in a few cases, the enzyme
activity slightly decreased. The highest decrease was observed for D7
at 120 mins of incubation period (−76.78%). Similarly, the enzyme
activity enhanced for majority of the cases when tap water detergent
formulations were tested for their compatibility to the test enzyme
preparation. The highest improvement was seen for D2 at incubation
of 120 mins (+48.8%) for tap water and D1 at 120 mins of incubation
period (+44.20%) for hard water samples. The results indicate the high
potential of the crude enzyme to be used in detergent formulations as
in majority of the cases increase of enzyme activity was seen.
For mutated strain (Table 2), for distilled water, the enzyme ac-
tivity was found to enhance in the presence of different detergents
for at least, one incubation time. This indicated that the detergent
formulations were somehow improving the enzyme activity. This may
due to the presence of surfactants in the detergent formulations. The
highest increase in enzyme activity was found to be for D2 (+19.40%)
at 20 mins of incubation period while highest loss in case of D6
(−56.60%) at 120 mins of incubation period. In the presence of tap
water, the enzyme activity was found to improve for 5 (D1–5) of the
7 detergent formulations used for at least one incubation period. For
D6 and D7, the enzyme activity decreased as the incubation period
increased. The highest improvement in enzyme activity was seen for D4
(+30.60%) at 120 mins of incubation time indicating high compatibility

3
S. Singh, J. Mangla and S. Singh
Fig. 1. Comparison of % enzyme activity for enzyme–detergent combinations of
Enzyme A and Enzyme B in (a) Distilled water at 60𝐨 C (b) Tap water at 60𝐨 C (c)
Tap water at 80𝐨 C (d) Hard water at 60𝐨 C.
of the enzyme with the said detergent formulation. For D6, a decrease
of 50.40% in enzyme activity at 120 mins of incubation time was
observed. This was the highest loss in enzyme activity in the presence
of tap water. Interestingly, even hard water+detergent formulations
were found to improve enzyme activity at different incubation period.
All the detergent formulations improved enzyme activity at least one
of the incubation periods tested in the present study. In contrast to
4
Resources, Environment and Sustainability 5 (2021) 100030
some of the previous studies (de Carvalho et al., 2008) where, at least,
some enzyme activity was lost in the presence of different detergents,
our study yields significant finding that the enzyme efficiency might
increase in the presence of detergents.
On comparing the stability of enzymes A and B, in the presence of
different detergents (Fig. 1), it was found that the Enzyme A, obtained
from wild-type A. fumigatus NTCC1222, was better compatible with the
detergents except for detergent D7 for distilled water, D4 for tap water
and slightly that for D1, D2 and D3 for hard water. Overall, Enzyme A
(EA) was more stable as compared to Enzyme B (EB) in the presence of
detergents. In the presence of distilled water at 80 min of incubation,
the best stability and efficiency of the enzyme A (EA) was found to
be in case of D1 (showing an increase of 31.90% in enzyme activity),
while in case of D4, an increase of 30.20% in the enzyme activity was
observed, in comparison to the control . In case of EB, D2 followed
by D1, were the most favorable for the enzyme activity but, at 20
min of incubation time. For tap water, D2 showed the best increase
in enzyme activity (48.80% increase in EA and 30.60% increase in
EB) at 120 min of incubation. For hard water, D1 was the most
preferable for both EA and EB. Overall, our crude amylase preparation
exhibits high compatibility with different detergent formulations which
strongly support its potential exploration in detergent industry, and
even appears to be better in performance at least as far as compatibility
with the detergent formulations is concerned (Dahiya et al., 2010).
The increase in amylase activity in the presence of detergents has
been reported earlier too (Sindhu et al., 2011; de Carvalho et al., 2008;
Yang et al., 2013). Though it is not very clearly known yet, the increase
in enzyme activity in our study might be due to enhancing influence of
surfactants already present in the commercial detergent formulations.
This might be due to ionic interactions and hydrophobic or hydrophilic
interactions from the fusion peptide (Sindhu et al., 2011; Siddiqui et al.,
2009). A subsequent decrease in enzyme activity beyond certain period
of incubation might be due to loss saturation effects beyond which, the
presence of surfactants might no longer enhance the enzyme activity.
The satisfactory sustenance of enzyme activity, even in the presence of
hard water (containing calcium or magnesium), might be due to the
fact that hard water inhibits the self-degeneration of enzymes. Further,
once the enzyme solution and detergent combine, enzyme effectiveness
remains substantially undiminished over long periods of time. Similar
observations have been made Marvin and Wiesenfeld (2014).
4. Conclusion
The current investigation explores the possible application of crude
amylase preparations obtained from different strains of indigenously
isolated fungus, A. fumigatus NTCC1222. As indicated, the enzyme
preparations were significantly compatible with all the detergent for-
mulations investigated, as indicated by the stability of the enzymes
preparations in specific incubation conditions. Interestingly, in some
cases, the enzyme activity was in fact, found to improve in the presence
of the detergent formulations under specific test conditions. The fact
that the enzymes were stable to specific, significant degree even in
the presence of poor quality (artificial hard water) water used further
enhances their use in detergent industry. The given study thus, adds
to the economic importance to the crude amylases of A. fumigatus
NTCC1222, and can surely be explored for use in detergent industries.
Declaration of competing interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared to
influence the work reported in this paper.
Acknowledgment
The first author is thankful to Lovely Professional University, Pun-
jab, India for providing basic financial and infrastructural facilities for
conducting the research and support at all levels.
S. Singh, J. Mangla and S. Singh
References
Akpan, I., Bankoley, M.O., Adesemowo, A.M., 1999. Production of 𝛼-amylase from
Aspergillus niger using cheap medium. Tropical Sci. 39, 77–79.
Anto, H., Trivedi, U., Patel, K., 2006. Alpha amylase production by Bacillus cereus MTCC
1305 using solid-state fermentation. Food Technol. Biotechnol. 44 (2), 241–245.
Balkan, B., Ertan, F., 2007. Production of a-amylase from Penicillium chrysogenum under
solid-state fermentation by using some agricultural by-products. Food Technol.
Biotechnol. 45 (4), 439–442.
Borchert, T.V., Lassen, S.F., Svendsen, A., Frantzen, H.B., 1995. Oxidation stable
amylases for detergents. Progress Biotechnol. 10, 175–179.
Dahiya, P., Kishore, V., Sheikh, S., Arora, P., 2010. Compatibility and wash performance
analysis of 𝛼-amylase from Bacillus amyloliquifaciens MTCC(610) with commer-
cial detergents. In: International Conference on Environmental Engineering and
Applications. pp. 160–164, http://dx.doi.org/10.1109/ICEEA.2010.5596114.
de Carvalho, R.V., Côrrea, T.L.R., da Silva, J.C.M., Mansur, L.R.C.de O., Martins, M.L.L.,
2008. Properties of an amylase from thermophilic Bacillus sp.. Braz. J. Microbiol.
39, 102–107.
de Souza, P.M., de Oliveira Magalhães, P., 2010. Application of microbial 𝛼-amylase
in industry- A review. Braz. J. Microbiol. 41 (4), 850–861.
Gurkok, S., 2019. Microbial enzymes in detergents: A review sumeyra GŨRKÕK. Int.
J. Sci. Engg. Res. 10 (9), 75–81.
Khajuria, R., Singh, S., 2020. Fungal amylase for detergent industry. In: Microbes
in Agricultural and Environmental Development. Academic Press. CRC-Taylor and
Francis Group, pp. 153–164.
Khambhaty, Y., 2020. Applications of enzymes in leather processing. Environ. Chem.
Lett. 18, 747–769.
Marvin, L.M., Wiesenfeld, A., 2014. Stable enzyme containing liquid detergent,
Publication number: US3697451 A. http://www.google.com/patents/US3697451.
Miller, G.L., 1959. Use of dinitrosalicyclic acid reagent for the determination of reducing
sugars. J. Anal. Chem. 31 (1959), 426–429.
Mitidieri, S., Martinelli, A.H.S., Schrank, A., Vainstein, M.H., 2006. Enzymatic detergent
formulation containing amylase from Aspergillus niger: A comparative study with
commercial detergent formulations. Biores. Technol. 97 (10), 1217–1224.
Pandey, A., Nigam, P., Soccol, C.R., Soccol, V.T., L. Vandenbegh, L., Mohan, R., 2000.
Biotechnological potential of agro-industrial residues II: Cassava baggase. Biores.
Technol. 74, 81–87.
Rodríguez, V.B., Alameda, E.J., Gallegos, J.F.M., Requena, A.R., López, A.I.G., 2006.
Thermal deactivation of a commercial 𝛼-amylase from Bacillus licheniformis used in
detergents. Biochem. Eng. J. 27 (3), 299–304.
Resources, Environment and Sustainability 5 (2021) 100030
Siddiqui, K.S., Parkin, D.M., Curmi, P.M.G., Francisci, D.D., Poljak, A., Barrow, K.,
Noble, M.H., Trewhella, J., Cavicchioli, R., 2009. A novel approach for enhancing
the catalytic efficiency of a protease at low temperature: reduction in substrate
inhibition by chemical modification. Biotechnol. Bioeng. 103, 676–686.
Sindhu, R., Suprabha, G.N., Shashidhar, S., 2011. Purification and characterization of a-
amylase from Penicillium janthinellum (NCIM 4960) and its application in detergent
industry. Biotechnol. Bioinf. Bioeng. 1 (1), 25–32.
Singh, S., 2016. Aspergillus enzymes for textile industries. In: Gupta, V.K. (Ed.),
New and Future Developments in Microbial Biotechnology and Bioengineering-
Aspergillus System Properties and Applications. Elsevier, Netherlands, pp.
191–199.
Singh, S., Cheema, S.K., Kaur, B., Mann, N.K., 2013a. Influence of ethanol on growth
alpha-amylase production for Aspergillus fumigatus NTCC1222 under solid state
fermentation. Int. J. Eng. Res. Technol. 6 (8), 67–69.
Singh, S., Cheema, S.K., Kaur, B., Mann, N.K., 2013b. Sorbitol: an enhancer of growth
and alpha-amylase production for Aspergillus fumigatus NTCC1222 using wheat bran
as substrate. Int. J. Biotechnol. Bioeng. Res. 4 (6), 555–560.
Singh, S., Cheema, S.K., Kaur, B., Mann, N.K., 2015. Effect of Ferrous sulphate on
growth and alpha-amylase production for Aspergillus fumigatus NTCC1222. Orient.
J. Chem. 31 (2), 1181–1184.
Singh, S., Dutt, D., Tyagi, C.H., Upadhyaya, J.S., 2010. Bio-conventional bleaching of
wheat straw soda-AQ pulp with crude xylanases from SH-1 NTCC-1163 and SH-2
NTCC-1164 strains of Coprinellus disseminatus to mitigate AOX generation. New
Biotechnol. 28 (1), 47–57.
Singh, S., Kaur, B., Mann, N.K., Cheema, S.K., 2013. Influence of calcium chloride on
growth and alpha-amylase production for wild and UV-mutated strains of Aspergillus
fumigatus. Int. J. Biotechnol. Bioeng. Res. 4 (7), 697–702.
Singh, S., Mann, N.K., Kaur, B., Cheema, S.K., 2013d. Effect of toluene on fungal growth
and amylase production- A step towards exploration of enzymes for industrial
applications. Int. Rev. Appl. Eng. Res. 4 (2), 117–122.
Singh, S., Singh, S., Mangla, J., 2016. Physical and chemical mutation for enhanced
alpha-amylase production by aspergillus fumigatus NTCC1222 under solid state
fermentation conditions using agri-residue waste. J. Pharm. Nutr. Sci. 6 (1), 22–26.
Singh, S., Singh, S., Sharma, L., Bali, V., Mangla, J., 2014. Production of fungal
amylases using cheap, readily available agri-residues, for potential application in
textile industry. Biomed. Res. Int. 215748, 9 pages, http://dx.doi.org/10.1155/
2014/215748.
Singh, S., Tyagi, C.H., Dutt, D., Upadhyaya, J.S., 2009. Production of high level of
cellulase-poor xylanases by wild strains of white rot fungus coprinellus disseminatus
in solid state fermentation. New Biotechnol. 26 (3–4), 165–170.
Yang, H., Liu, L., Shin, H.D., Chen, R.R., Li, J., Du, G., Chend, J., 2013. Integrating
terminal truncation and oligopeptide fusion for a novel protein engineering strategy
to improve specific activity and catalytic efficiency: Alkaline 𝛼-amylase as a case
study. Appl. Env. Microbiol. 79 (20), 6429–6438.