Amylase assay was done by using a reaction mixture consisting

1 ml of substrate solution (1.1% soluble starch in 50 mM phosphate

buffer pH 7.2) and 100 μl of the enzyme solution. The reaction

mixture was incubated for 10 min at 30oC. Reaction was stopped by

adding 2 ml of dinitrosalicyclic acid (DNSA) reagent .

The reaction mixture was heated to 100oC for 10 min and cooled.

Optical density of each sample with reaction mixture was taken at

575 nm in a spectrophotometer (Shimadzu, Japan). Enzyme activity

was expressed in units (1 unit/ml = amount of enzyme which

releases 1 μ mole glucose under the assay condition).

http://ijpbs.com/ijpbsadmin/upload/ijpbs_5387f2cb9c9a2.pdf

https://www.google.iq/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&cad=rja&uact=8&ved=0ahUKEwjkr8bGyJXTAhXLtBQKHUTBAt4QFggdMAA&url=http%3A%2F%2Fwww.ajol.info%2Findex.php%2Fajb%2Farticle%2Fdownload%2F59286%2F47584&usg=AFQjCNFR1VIHHmQi0i_D04BRq4kGT8mOQw&sig2=haBV9E4D2DQGEJqvIv1PKQ

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3209940/

you can go to link for protocol

https://www.researchgate.net/profile/Tabinda_Salman/publication/298807760_Medium_optimization_for_the_production_of_amylase_by_Bacillus_subtilis_RM16_in_Shake-flask_fermentation/links/56ec5a5b08ae4b8b5e7334a4/Medium-optimization-for-the-production-of-amylase-by-Bacillus-subtilis-RM16-in-Shake-flask-fermentation.pdf

Article Medium optimization for the production of amylase by Bacillu...

112

Hi Lala

You can read some references in attachment 1

https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/a3403enz.pdf

2

https://ps.oxfordjournals.org/content/92/9/2389.full

3

https://www.mdpi.com/2311-5637/3/1/7/htm

4

https://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822000000400011

5

https://secure.megazyme.com/Alpha-Amylase-Bacillus-licheniformis

6

http://www.academicjournals.org/article/article1380883954_Nimrat%20and%20Vuthiphandchai.pdf

7

http://www.tandfonline.com/doi/abs/10.3109/09637486.2013.832175?scroll=top&needAccess=true&journalCode=iijf20

8

https://worldwidescience.org/topicpages/b/bacillus+licheniformis+alpha-amylase.html

9

http://scholar.sun.ac.za/bitstream/handle/10019.1/96885/hunter_probiotics_2015.pdf?sequence=2

10

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0144804

11

https://www.animalsciencepublications.org/publications/jas/articles/89/6/1795

12

https://www.trjfas.org/pdf.php?id=194

13

https://www.science.gov/topicpages/b/bacillus+licheniformis+alpha-amylase.html

14

https://www.ijrbat.in/upload_papers/28012015112856Paper%20_4_.pdf

15

https://journal.frontiersin.org/article/10.3389/fmicb.2017.00514/pdf

16

http://www.microbiologyresearch.org/docserver/fulltext/micro/143/8/mic-143-8-2733.pdf?expires=1491851813&id=id&accname=guest&checksum=F6D9B7EC2B05D82394FF807251C09109

17

https://brage.bibsys.no/xmlui/bitstream/id/204311/Kristian%20Kollerud.pdf

18

https://brage.bibsys.no/xmlui/bitstream/id/204311/Kristian%20Kollerud.pdf

19

https://`aut.researchgateway.ac.nz/bitstream/handle/10292/4606/HadiJA.pdf?sequence=4

Good Luck