<110>	Algenuity Holdings Ltd
<120>	MODIFIED STRAINS OF CHLORELLA MICROALGAE SPECIES HAVING REDUCED CHITIN CONTENT 
<150> 	GB2007940.6
<151>	27/05/2020
<160>	3
<210>	1
<211>	744
<212>	PRT
<213>	Chlorella Vulgaris
<222>	347
<223>	Mutation in an ABC transporter (Arg347Cys) of a modified strain
<400>	1

Met Trp Ser Ala Gly Leu Gln Phe Gln Ala Ser Ala Thr Lys Leu Pro
                  5		     10	                 15
Ser Arg Arg Arg Pro Leu Pro Ala Ala Arg Pro Pro Cys Leu Val Ala
	    20		        25		     30	
Lys Arg His Ala Ala Ala Gly Ala Val Arg Gln Gln Gln Gln Arg Trp 
	35		    40   	         45	
Pro Trp Arg Thr Ile Pro Arg Ala Ala Ala Leu Ala Ala Ala Ala Ala
     50			55		     60			
Ala Ala Ala Asp Asp Pro Ser Ala Thr Gln Pro Ala Leu Ile Asp Gly
65		     70		         75		     80
Asp Asp Ala Ala Ala Ala Ala Thr Gly Gly Val Ala Ala Ser Thr Ala
		 85	             90		         95
Leu Ala Pro Thr Pro Ala Ala Ala Ala Asp Val Gly Leu Ser Leu Ser
	    100		        105	            110
Glu Leu Trp Gln Leu Leu Gln Gln Asp Arg Arg Arg Leu Ala Ala Cys
	115	            120	        	125
Ile Ala Cys Thr Ala Val Ser Val Ala Ser Ala Val Leu Val Ala Pro
    130		        135		    140
Cys Leu Gly Arg Val Val Asp Ile Ile Ser Arg Gly Thr Ala Ala Thr
145		    150	        	155       	    160
Pro Arg Glu Leu Ala Val Ala Val Gly Arg Leu Gly Thr Val Tyr Val
                165		    170		        175
Ile Ala Asn Val Gly Met Ala Val Gln Val Ala Leu Ser Leu Ala Leu
	    180		        185	            190	
Gly Glu Gly Leu Ala His Arg Leu Arg Cys Arg Leu Phe Gly Ala Leu
        195	            200		        205	
Leu Ser Arg Asp Thr Leu Phe Phe Asp Gln Val Lys Thr Gly Gln Met
    210			215	            220
Val Ala Trp Leu Gly Gln Asp Val Glu Val Leu Gln Ser Thr Val Ser
225		    230		        235		    240	
Lys Leu Leu Gly Ala Arg Gly Ile Arg Ser Ala Phe Glu Thr Cys Gly
	        245		    250		        255	
Ile Ile Ile Val Leu Phe Thr Leu Ser Trp Pro Leu Ala Ala Ala Leu
	    260		        265	            270
Leu Val Ser Ala Pro Leu Leu Thr Pro Leu Ile Ala Arg Leu Ser Ser
        275		    280		        285
Ala Ile Gly Ile Ala Ser Lys Ala Ser Gln Ala Ala Ser Ser Glu Val
    290		        295		    300     
Ser Ala Ala Ala Asp Glu Val Val Glu Asn Met Arg Val Val Lys Leu
305		    310	                315		    320	
Phe Ala Gln Gln Gln Ala Glu Leu Lys Arg Phe Gln Gly Leu Leu Gly
	        325		    330		        335
Thr Ala His Gln Leu Ala Leu Lys Val Leu Cys Leu Gln Ala Leu Leu
	    340		        345		    350		
Asp Gly Ser Ser Arg Val Arg Asn Thr Leu Cys Val Leu Ala Thr Leu
        355		    360		        365			
Gly Leu Gly Ala Tyr Met Ala Leu His Ser Ala Val Ser Ile Gly Thr
    370		        375	            380			
Cys Tyr Ser Phe Phe Val Phe Ser Phe Ser Phe Ala Phe Ala Leu Gly
385		    390		        395		    400		
Asn Leu Thr Asn Thr Val Gly Asp Val Ala Arg Ala Ala Gly Ala Ile
                405		    410		        415
Asn Arg Ala Met Arg Thr Met Gln Gln Ala Leu Gly Thr Ala Ser Thr
   	    420		        425	            430
Glu Ala Thr Ala Ala Leu Leu Ser Ser Ala Asp Asn Ala Pro Leu Glu
        435		    440		        445	
Ala Gly Gly Asp Ala Val Thr Gly Ala Gln His Gly Arg Gln Leu Pro
    450		        455		    460
Ala Gly Trp Arg Gly Glu Ile Glu Phe Gln Asn Val Gly Phe Ser His
465		    470		        475		    480
Pro Gly Gly Trp Ala Ile Lys Asp Leu Ser Phe Lys Leu Pro Pro Gly
	        485		    490		        495	
Ser Thr Val Ala Leu Val Gly Pro Ser Gly Gly Gly Lys Thr Thr Ile
            500	                505	            510	
Ala Ser Met Leu Met Lys Leu Tyr Asp Val His Ala Gly His Ile Leu
        515		    520		        525
Val Asp Gly Val Pro Leu Asp Gln Leu Asp Thr Lys Trp Trp Arg Gln
    530		        535	            540
Gln Leu Gly Val Val Met Gln Ser Pro Gly Leu Leu Thr Gly Arg Ile
545		    550		        555		    560	
Ala Asp Ile Ile Arg Tyr Gly Gln Pro Gly Ala Ser Asp Ala Asp Val
	        565		    570		        575
Ala Arg Ala Ala Arg Ala Ala Gln Ala Asp Gly Phe Ile Glu Ala Leu
	    580		        585		    590	
Pro Asn Gly Tyr Gln Thr Val Ile Gly Ser Gly Ser Gly Ile Glu Leu
        595		    600		        605	
Ser Gly Gly Gln Gln Gln Arg Leu Ala Ile Ala Arg Ala Leu Leu Pro
    610   	        615		    620
Arg Pro Arg Leu Leu Ile Phe Asp Glu Ala Thr Ser Ala Leu Asp Val
625		    630		        635		    640 	
Glu Thr Glu Gln Gly Val Thr Gln Ala Leu Glu Gly Ala Gly Arg Gly
	        645		    650		        655
Val Thr Ser Leu Val Ile Ala His Arg Leu Ser Thr Val Arg Arg Ala
	    660		        665		    670			  	
Asp Leu Ile Val Val Val Ala Ser Gly Lys Val Val Glu Gln Gly Thr
        675		    680		        685
His Glu Gln Leu Met Gln Arg Gln Asn Gly Val Tyr Trp Ser Leu Val
    690		        695		    700
Ser Gly Ala Glu Ser Arg Gly Gln Asp Ser Trp Glu Asp Asp Asp Glu
705		    710		        715		    720	
Asp Asp Glu Glu Gln Asp Val Gln Gly Glu Gly Lys Gln Pro Pro Met
	        725		    730		        735			  
Ala His Glu Leu Ala Pro Thr Ala
	    740

<210>	2
<211>	5313
<212>	DNA
<213>	Chlorella Vulgaris
<222>	2750
<223>	Mutation localised at a splicing donor site after exon 9 at the end of the WD40 repeat domain at a position 2750 of a gene encoding for a guanine nucleotide-binding protein in modified strain
<400>	2

atggcctccg ctggcggcgc ggtgccgcca cgcaaagtgt cgttcctctt tcggcgcgag 60
ggcatagggg tgggtacttg caggagtgca gaggttgtgc ttggcggtgg cttacagtca 120
aacgcagaag gacagcgtcg ctcatctggt tgccccggtc cccatgcctc ccccttgccg 180
ccgcatcaat gttcctgcac cctcccctcc catcctcacg cagcactgcg gaggggtgaa 240
tgccctggct ttatccaggg acgaacagca gctgtggacc ggttccaggg actcaaccat 300
ctgctggtgc gaggcagctg cgctggccag cagtacgctg cgcttggtcg caaccctcct 360
ttgccagtcg ggtgccagca gcattttcgc ctctcagctt caagctacag cgtcatgttc 420
tcagtcacgt catgtgtcct gcctgcagct ggtccacagc tgagaaagcg cagccggatc 480
tgatgcacga gcacgtcggt cacattgact gggtgggtgt tgcaggcgcc ctcgtcacac 540
cctcgtgtgc ttgtgcatgt atgctcaaat gtcgcgttgg atcatcacag cgttgtgtgg 600
ttgtaagggg tctcatgggc acctgcactg ctcttcattg gagctcagca gggcttcatg 660
ctgcgtggtg ttgatgccag ctctctccaa gcagccccag caaaccagaa gatgcaccac 720
cctctgcccc agtgctgacc tccagcctgc tgtgccgtga tgtgtccgca ggtcaatggg 780
ctggcactgg caggcgatgt gctggtgtcc tgctcctccg acaaaacgct gcgggtgtgg 840
cagcccgact cgccaggtca gcagcacgtg ccagctcagc tttggctgtg ctggccgcct 900
ctgcaactgc tgcaggtgtt tgctctccgc ttttgccaaa ggagtctcgc ctgccccacc 960
ctggcccaac ctagtcgctc acccacccca cccgcttcac actttgtgtc cccccccagc 1020
agagccagcg ctgggctgcc tgcagggaca cagcgactat gtcaccgcgc tggctgcgtg 1080
cgccgacggc ggcgtgctgg cctcgggcgg cctgcgtggc gaggtgctgc tgtgggacct 1140
ggccacgctg cggggcatca tggcgggcag cacccaggtg aggctttgag tgctgccagc 1200
tgtcctgtgc tgccgacctg cagctgctgc ccacctgcag ctgctacatg caccggctgc 1260
actgatatgg ctctcttgat tacagacaat gcacaatcat acacctgtaa gacaattgtg 1320
gtcataaatc atgtgcgcag gaccaaggtt acagcgcatc aacagcggag gtgccccgtg 1380
cttcagtgta tgctgtggcc atcaatgctg ctggcacgct gattgcggca gggtcgaccc 1440
aggtgcggca cgacctgggg gctggacagt gcagcagtgt cgaatggcag ctgcagagct 1500
gaaggatgca gactcggctt gcaggctgca cgtgtgacga ttctcacggc cgacactgct 1560
gtctgttgcg atgtttccga tgtgctcctg cctgcacatt cgacctttgt acttctccgg 1620
cttgcttgat gcgatgtgct gtgcagggcg tgagcgtggt ggacgtccgc agcgggcgca 1680
gcgtgatgca gctcaagggc cacacagaca acgtcaggtg tgtgtgtgtg gggggggggg 1740
ggggtggcgg tggggagtgt gtggcgcagt gtgtgcgttg gagtgttctg ttgctgcttc 1800
ctttccaaag cgcttgcagc tctgggtgat ggctgcccct tctgtggtct cagcacaccc 1860
tttcctcaca caggtgccgt gcgcctggtg ttgcctggtg cacacctgtt gtttttcccg 1920
aaaccttggg gtgccattcc aattccgagc gtgctgcacc gctcaacctc accccaccac 1980
cacccagcac cgcattggtg ccgactgctc taatcgtggt tgttgttgaa tcactgcctg 2040
tctgcaggtc gctgcagctt gaagcagagg ggcggctgct tctgagcggg tctgcggacc 2100
atacaatgcg gctgtgggac ctgggccagc agcgctgtgt gcagacgctg gccgtgcaca 2160
ctgacagcgt gtgggcgctg gcggccaccc cctcctttgc ggcggtgttc agcggcgggc 2220
gggacggctg catataccgg tgggtgcatt cagatgtgtg tgtgtgtgtg tctgtggtgg 2280
tggtggggag tgtggtggtg cggccggcgg ctgtgccagt ggggacaccc tgtgacggca 2340
tgaccttgcg ggttccctct ctgcctcttg cccaccatgg ctgctgaccc atcatgtcac 2400
tgcctccctg tcactccacc accctgcagc acccgcctgt cgacccgggt tagcgagctg 2460
ctggctgtgg agcagcggcc ggtgctggcg ctggctgtgg cctcggacga gcggcggctg 2520
tgggcagcca ccactgcgcc cagcgtgcac acctggcacc tggagttggc tgcagcagct 2580
tcggcacaca ccgcggcgat aggcggcggc agcaccagcg ttggcggcgg cacgccgcgg 2640
tcgccgctgg gggctgccgc cggggctgcc gccgtgctgg gcggtccctg cttcattgcc 2700
agcccctctg cagccgcccg agcacgacta ccgtttgctg gacagcagga tgaggcactg 2760
ttgctgctga ctgcgctgcc tgctgctgct atgctgctct gattgcaaat ggtgccgccg 2820
ctggtccagc acctgtctcc catcacgcct tccagtgtcc tgtttgattc gccgccattg 2880
taatgcaaca gaccgccagc catcgtcctt gctgcctgcc ggcctgcagg ctcccatcca 2940
ccgccgcagc agggcgagcc agccgccgcc acgcccggtc tgccgccaat tcagcatgcc 3000
gtggtgctca ctgacaggcg gcatgtgctg acccgagatg cggatggcac agtgctgtgc 3060
tgggacctca cagcaggtga gatggggtgt gcagtgcttg ggcggcaggc actcacacag 3120
ctgacagaag ttgctggata cactgtcgca cgtgcagcct acgtcgcttt atcaaacaat 3180
ctgcattgta tccaaccaga caatagaatc tgcaaccgta tcccacccca cacgccccgc 3240
cctcactcac cctgcaggct gtgtgacccg cgactatggt cgctgcgagt tgaaagaggt 3300
ggaggcggca ctcttccagc ccaaccactc tgtatctgcc tggtgccagc ccgaggtcaa 3360
gctgggcagc ctggggggtg ccatggagcc accgcagtgc tttggtgctg agatctatgc 3420
ccaggtgcgg cagttgtttc cagatactca tgtcacactg tttcaagctt aagcgctgca 3480
ggctggtgca caaattccac tcattacacg acttctgttc aggatgccgc cccatgtttc 3540
ttttgctcca gatatcactt tgcgtgtgaa tgagcaagtc tggcgttgcc atgcctagct 3600
cgccctgctc ctgctttctc ctcgtgtcat gccacctcca tccgcgtcgt tgcacctgca 3660
ggacctggga gacgaggagg cacctgtcga cttgaaagtc aactttgggg agcagatgct 3720
ggtggcttta ttcaagggct gggcagaagc ggcgcagcgc agcagctgcc acgctctgcc 3780
tgctccctcc acgcaagttc tgcggccagc acaagagcgg gtcctgggca gccgggcagt 3840
agaccagcag tctggtgctg agccaagcga cagcggcggc agccacggcg gcgatggcga 3900
gcggcaaagc aatgtggcgg cggagcaggc ggcagcgggg gggtgcgaac gcagcaccct 3960
gcaggatggt gacagtgagg ggcagagtat tgacgggtga gcggcagcat gcaccgcact 4020
tgcgcagcta tagtcagcga tggatggaag catagtcatg acagagctgt gggtgtaggc 4080
acctgcaaga tggaggcttt ctttctgaat ttcgggccgg cggtgttcag gcttggccgg 4140
ccatcctgct cttgtcctgg cagcaagtac tcgcaaacct gcacgtgtgt gtgagtgctg 4200
cggcgtcctg ctcgtgcact ggtcgctcac cctgccagcc tgccctcctc gccacacagg 4260
gagcccgacc aagccactcg cttcaagttt ctcaccccgc tggcacctgt ggtgatggtc 4320
agcggcagtg gcggcagcgg taccagagct ggcggtaccg tgccctggag gtgccctact 4380
gatgcgttca ccgggaagga agacgtgcct caggtgctgt ccggggtcgt gtcaggaagg 4440
aagctgcggc tgggggcccg acagcaccag aggcaaccag agcacttggc atttgcacgc 4500
ctttcgacgc tctggtgtag ctgttgactc tggcccgctt gctgcctcct gccctgctct 4560
gccgcagtgg gtagccgact gcgtcctacg gggccggtat ccggtcagca gggagctcaa 4620
gatggcgttc cagctgatgc ccctgccggt aagcccttgc atacaagtag tggggctgat 4680
aactgccagc ttgcacgatc cagcagctga gaacagtgaa gcgctttgca gcgcaaatgc 4740
tactgtggcg cagtgccaag ccgtcgtgac tcatctcaaa cccctctctc aaccgccctc 4800
actatacacc gtttgcctgc ctgcagggct cgccgctgcc atcgctgctg caaagcaagc 4860
tgaatgcacc gagggtgctg cgggtcaaca aggtggctga ctacatccgc cgcaagatgg 4920
aggagcaggg catagcattg cgggaggagc cgctgttttg ggatggcagc aaggaggaga 4980
agtggcacgc ggagcaagct gccagtgccg acggcagcaa cgacggcagc agtgggggcc 5040
ttgtctcact gccggtgcac cagctgcacc ctgtgcagcc gcttccgggc gccgccagca 5100
gcatcagccc cgcggagagg gcccggggca gcggtttggc cgggcacgga gcgcttttgg 5160
tcacgtgtgc gggcaacact gtgccttggg actttagctt ggcagcagta cgccagtgga 5220
tttggcggcg ctctgacgat ttggtgctgc attactctgt gagggatcgc tccacgccgc 5280
tgaagctgcc agccattcga cgtccttcat gaa 		   	          5313

<210>	3
<211>	473
<212>	PRT
<213>	Chlorella Vulgaris
<222>	46
<223>	Mutation in an aquaporin (Ala46Val) of a modified strain
<400>	3

Met Ser Gly Gly Thr Ala Val Asn Asn Thr Tyr Ser Leu Ala Ala Glu 
                  5		     10		         15
Cys Gly Ser Glu Met Ile Gly Thr Cys Phe Val Ile Tyr Leu Gly Glu 
	    20		         25	             30	
Ser Ile Ile Ala Asn Glu Leu Leu Ser Lys Thr Lys Gly Val Arg Met 
	 35		     40		         45	
Gly Trp Leu Ala Val Ala Leu Gly Phe Gly Leu Ser Phe Gly Val Val 
     50		         55		     60			
Ile Met Met Leu Gly Tyr Ile Ser Ala His Leu Asn Pro Ala Thr Cys 
65		     70		         75		     80
Leu Ala Leu Trp Val Ile Gly Lys Ile Pro Phe Thr His Phe Leu Ala 
		 85		     90		         95
Leu Ala Gly Ala Glu Phe Ala Gly Ala Phe Val Gly Ala Cys Leu Val 
	    100		        105	            110
Phe Leu His Tyr Ile Pro His Phe Lys Thr Val Pro Glu Pro Met Pro 
	115	            120		        125
Arg Asn Glu Asp Asp Val Leu Leu Arg Ser Arg Asp Ala Leu Thr Phe 
    130		        135		    140
Glu Ala Leu Asn Ile Ala Ser Tyr Asp Thr Arg Ala Arg Gln Arg Thr 
145		    150		        155		    160
Ala Ser Ser Lys Gly Gly Val Met Gly Val Ile Asn Asp Ala Ile Lys 
	        165		    170		        175
Asp Val Arg Tyr Tyr Phe Lys Glu Thr His Gln Ala Pro Gln Glu His 
	    180		        185		    190	
Val Glu Leu Val Glu Val Ala Leu Gly Pro Ala Glu Met Lys Gly Ala 
        195	            200		        205	
Val Glu Asn Arg Leu Arg Arg Arg Ser Val Gln Val Cys Asp Val His 
    210		        215	            220
Arg Arg Leu Lys Asp Met Ser Ile Glu Glu Phe Lys Glu Lys Leu Gln 
225		    230		        235		    240	
Val Lys Ser Ser Ala Leu Asn Gly Gly Met Pro Arg Ser Ala Ser Ile 
	        245		    250		        255	
Asn Leu Gly Glu Leu Asp Arg Ala Met Gly Gly Glu Gln Gly Ala Ala 
	    260		        265		    270
Glu Gly Lys Asp Leu Gly Asp Ser Ser Ser Gly Glu Glu Ala His Gln 
        275		    280		        285
Gln Arg Val Gln Arg Gln Ala Thr Ala Pro Pro Gln Val Gln Gly Pro 
    290		        295		    300      
Pro Thr Ala Trp Lys Asp Lys Leu Met Arg His Ile Glu Arg Tyr Gly 
305		    310		        315		    320
Glu Arg Gln Ala Arg Val Phe Asp Ala Ala Val Ile Ala Asp Gln Asn 
	        325		    330		        335
Ala Lys Leu Ser Ile Phe Cys Thr Arg Pro Ala Ile Tyr Ala Pro Val 
	    340		        345		    350		
Phe Asn Phe Leu Thr Glu Val Met Cys Thr Thr Ala Leu Val Phe Gly 
        355		    360		        365			
Ala Leu Met Met Tyr Ala Arg Arg Asp Leu Leu Asn Pro Glu His Lys 
    370		        375	            380			
Ala Leu Phe Gln Ser Tyr Glu Gly Met Trp Ile Gly Phe Phe Val Phe 
385		    390		        395		    400
Val Ala Ile Leu Gly Leu Gly Gly Pro Thr Gly Ile Ala Ala Asn Pro 
	        405		    410		        415
Ala Arg Asp Phe Ser Pro Arg Leu Ala His Ala Leu Leu Pro Ile Ala 
	    420		        425		    430
Gly Lys Gly Pro Ser Glu Trp His Tyr Gly Trp Ile Pro Phe Trp Ala 
        435	 	    440		        445	
Pro Phe Phe Gly Gly Ala Ala Ala Gly Gly Leu Tyr Leu Leu Val Gln 
    450		        455		    460
Met Leu Asn His Ser Lys Tyr Val Ala 
465		    470



